Prmt5 inhibitors and uses thereof

ABSTRACT

Described herein are compounds of Formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof. Compounds of the present invention are useful for inhibiting PRMT5 activity. Methods of using the compounds for treating PRMT5-mediated disorders are also described.

RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(e) to U.S.provisional patent application, U.S. Ser. No. 61/790,928, filed Mar. 15,2013, and to U.S. provisional patent application, U.S. Ser. No.61/745,490, filed Dec. 21, 2012, the entire contents of each of whichare incorporated herein by reference.

BACKGROUND OF THE INVENTION

Epigenetic regulation of gene expression is an important biologicaldeterminant of protein production and cellular differentiation and playsa significant pathogenic role in a number of human diseases.

Epigenetic regulation involves heritable modification of geneticmaterial without changing its nucleotide sequence. Typically, epigeneticregulation is mediated by selective and reversible modification (e.g.,methylation) of DNA and proteins (e.g., histones) that control theconformational transition between transcriptionally active and inactivestates of chromatin. These covalent modifications can be controlled byenzymes such as methyltransferases (e.g., PRMT5), many of which areassociated with specific genetic alterations that can cause humandisease.

Disease-associated chromatin-modifying enzymes (e.g., PRMT5) play a rolein diseases such as proliferative disorders, metabolic disorders, andblood disorders. Thus, there is a need for the development of smallmolecules that are capable of inhibiting the activity of PRMT5.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Protein arginine methyltransferase 5 (PRMT5) catalyzes the addition oftwo methyl groups to the two ω-guanidino nitrogen atoms of arginine,resulting in ω-NG, N′G symmetric dimethylation of arginine (sDMA) of thetarget protein. PRMT5 functions in the nucleus as well as in thecytoplasm, and its substrates include histones, spliceosomal proteins,transcription factors (See e.g., Sun et al., 20011, PNAS 108:20538-20543). PRMT5 generally functions as part of a molecule weightprotein complex. While the protein complexes of PRMT5 can have a varietyof components, they generally include the protein MEP50 (methylosomeprotein 50). In addition, PRMT5 acts in conjunction with cofactor SAM(S-adenosyl methionine).

PRMT5 is an attractive target for modulation given its role in theregulation of diverse biological processes. It has now been found thatcompounds described herein, and pharmaceutically acceptable salts andcompositions thereof, are effective as inhibitors of PRMT5. Suchcompounds have the general Formula (I):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R⁸, R⁹, R¹⁰, R¹¹, R^(x), n, X, and Cy are as defined herein.

In some embodiments, pharmaceutical compositions are provided whichcomprise a compound described herein (e.g., a compound of Formula (I)),or a pharmaceutically acceptable salt thereof, and optionally apharmaceutically acceptable excipient.

In certain embodiments, compounds described herein inhibit activity ofPRMT5. In certain embodiments, methods of inhibiting PRMT5 are providedwhich comprise contacting PRMT5 with an effective amount of a compoundof Formula (I), or a pharmaceutically acceptable salt thereof. The PRMT5may be purified or crude, and may be present in a cell, tissue, or asubject. Thus, such methods encompass inhibition of PRMT5 activity bothin vitro and in vivo. In certain embodiments, the PRMT5 is wild-typePRMT5. In certain embodiments, the PRMT5 is overexpressed. In certainembodiments, the PRMT5 is a mutant. In certain embodiments, the PRMT5 isin a cell. In certain embodiments, the PRMT5 is in an animal, e.g., ahuman. In some embodiments, the PRMT5 is in a subject that issusceptible to normal levels of PRMT5 activity due to one or moremutations associated with a PRMT5 substrate. In some embodiments, thePRMT5 is in a subject known or identified as having abnormal PRMT5activity (e.g., overexpression). In some embodiments, a providedcompound is selective for PRMT5 over other methyltransferases. Incertain embodiments, a provided compound is at least about 10-foldselective, at least about 20-fold selective, at least about 30-foldselective, at least about 40-fold selective, at least about 50-foldselective, at least about 60-fold selective, at least about 70-foldselective, at least about 80-fold selective, at least about 90-foldselective, or at least about 100-fold selective relative to one or moreother methyltransferases.

In certain embodiments, methods of altering gene expression in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the cell in culture in vitro. In certain embodiments, cellis in an animal, e.g., a human.

In certain embodiments, methods of altering transcription in a cell areprovided which comprise contacting a cell with an effective amount of acompound of Formula (I), or a pharmaceutically acceptable salt thereof,or a pharmaceutical composition thereof. In certain embodiments, thecell in culture in vitro. In certain embodiments, the cell is in ananimal, e.g., a human.

In some embodiments, methods of treating a PRMT5-mediated disorder areprovided which comprise administering to a subject suffering from aPRMT5-mediated disorder an effective amount of a compound describedherein (e.g., a compound of Formula (I)), or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition thereof. Incertain embodiments, the PRMT5-mediated disorder is a proliferativedisorder, a metabolic disorder, or a blood disorder. In certainembodiments, compounds described herein are useful for treating cancer.In certain embodiments, compounds described herein are useful fortreating hematopoietic cancer, lung cancer, prostate cancer, melanoma,or pancreatic cancer. In certain embodiments, compounds described hereinare useful for treating a hemoglobinopathy. In certain embodiments,compounds described herein are useful for treating sickle cell anemia.In certain embodiments, compounds described herein are useful fortreating diabetes or obesity.

Compounds described herein are also useful for the study of PRMT5 inbiological and pathological phenomena, the study of intracellular signaltransduction pathways mediated by PRMT5, and the comparative evaluationof new PRMT5 inhibitors.

This application refers to various issued patent, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., a inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various isomeric forms, e.g., enantiomers and/ordiastereomers. For example, the compounds described herein can be in theform of an individual enantiomer, diastereomer or geometric isomer, orcan be in the form of a mixture of stereoisomers, including racemicmixtures and mixtures enriched in one or more stereoisomer. Isomers canbe isolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred isomers canbe prepared by asymmetric syntheses. See, for example, Jacques et al.,Enantiomers, Racemates and Resolutions (Wiley Interscience, New York,1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistryof Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofany compound described herein does not exclude any tautomer form.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of hydrogen by deuterium ortritium, replacement of ¹⁹F with ¹⁸F, or the replacement of a carbon bya ¹³C- or ¹⁴C-enriched carbon are within the scope of the disclosure.Such compounds are useful, for example, as analytical tools or probes inbiological assays.

The term “aliphatic,” as used herein, includes both saturated andunsaturated, nonaromatic, straight chain (i.e., unbranched), branched,acyclic, and cyclic (i.e., carbocyclic) hydrocarbons. In someembodiments, an aliphatic group is optionally substituted with one ormore functional groups. As will be appreciated by one of ordinary skillin the art, “aliphatic” is intended herein to include alkyl, alkenyl,alkynyl, cycloalkyl, and cycloalkenyl moieties.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 10 carbon atoms (“C₁₋₁₀alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms(“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbonatoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1 to 7carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl group has 1to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, an alkyl grouphas 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments, an alkylgroup has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In some embodiments, analkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). In some embodiments,an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”). In someembodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). In someembodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. In certain embodiments, each instance of an alkyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, thealkyl group is substituted C₁₋₁₀ alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens.

“Perhaloalkyl” is a substituted alkyl group as defined herein whereinall of the hydrogen atoms are independently replaced by a halogen, e.g.,fluoro, bromo, chloro, or iodo. In some embodiments, the alkyl moietyhas 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In some embodiments, thealkyl moiety has 1 to 6 carbon atoms (“C₁₋₆ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 4 carbon atoms (“C₁₋₄perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 3 carbonatoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In some embodiments, all of thehydrogen atoms are replaced with fluoro. In some embodiments, all of thehydrogen atoms are replaced with chloro. Examples of perhaloalkyl groupsinclude —CF₃, —CF₂CF₃, —CF₂CF₂CF₃, —CCl₃, —CFCl₂, —CF₂Cl, and the like.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon double bonds, and no triple bonds (“C₂₋₂₀ alkenyl”). Insome embodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms(“C₂₋₉ alkenyl”). In some embodiments, an alkenyl group has 2 to 8carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, an alkenyl group has2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In some embodiments, an alkenylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”). In some embodiments, analkenyl group has 2 to 5 carbon atoms (“C₂₋₅ alkenyl”). In someembodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂₋₄ alkenyl”).In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂₋₃alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂alkenyl”). The one or more carbon-carbon double bonds can be internal(such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples ofC₂₋₄ alkenyl groups include ethenyl (C₂), 1-propenyl (C₃), 2-propenyl(C₃), 1-butenyl (C₄), 2-butenyl (C₄), butadienyl (C₄), and the like.Examples of C₂₋₆ alkenyl groups include the aforementioned C₂₋₄ alkenylgroups as well as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and thelike. Additional examples of alkenyl include heptenyl (C₇), octenyl(C₈), octatrienyl (C₈), and the like. In certain embodiments, eachinstance of an alkenyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkenyl”) or substituted (a“substituted alkenyl”) with one or more substituents. In certainembodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl. Incertain embodiments, the alkenyl group is substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon triple bonds, and optionally one or more double bonds(“C₂₋₂₀ alkynyl”). In some embodiments, an alkynyl group has 2 to 10carbon atoms (“C₂₋₁₀ alkynyl”). In some embodiments, an alkynyl grouphas 2 to 9 carbon atoms (“C₂₋₉ alkynyl”). In some embodiments, analkynyl group has 2 to 8 carbon atoms (“C₂₋₈ alkynyl”). In someembodiments, an alkynyl group has 2 to 7 carbon atoms (“C₂₋₇ alkynyl”).In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C₂₋₆alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms(“C₂₋₅ alkynyl”). In some embodiments, an alkynyl group has 2 to 4carbon atoms (“C₂₋₄ alkynyl”). In some embodiments, an alkynyl group has2 to 3 carbon atoms (“C₂₋₃ alkynyl”). In some embodiments, an alkynylgroup has 2 carbon atoms (“C₂ alkynyl”). The one or more carbon-carbontriple bonds can be internal (such as in 2-butynyl) or terminal (such asin 1-butynyl). Examples of C₂₋₄ alkynyl groups include, withoutlimitation, ethynyl (C₂), 1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl(C₄), 2-butynyl (C₄), and the like. Examples of C₂₋₆ alkenyl groupsinclude the aforementioned C₂₋₄ alkynyl groups as well as pentynyl (C₅),hexynyl (C₆), and the like. Additional examples of alkynyl includeheptynyl (C₇), octynyl (C₈), and the like. In certain embodiments, eachinstance of an alkynyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents. In certainembodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl. Incertain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

“Carbocyclyl” or “carbocyclic” refers to a radical of a non-aromaticcyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C₃₋₁₀carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. Insome embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms(“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to6 ring carbon atoms (“C₃₋₆ carbocyclyl”). In some embodiments, acarbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆ carbocyclyl”). Insome embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms(“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groups include,without limitation, cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl(C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like.Exemplary C₃₋₈ carbocyclyl groups include, without limitation, theaforementioned C₃₋₆ carbocyclyl groups as well as cycloheptyl (C₇),cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇),cyclooctyl (C₈), cyclooctenyl (C₈), bicyclo[2.2.1]heptanyl (C₇),bicyclo[2.2.2]octanyl (C₈), and the like. Exemplary C₃₋₁₀ carbocyclylgroups include, without limitation, the aforementioned C₃₋₈ carbocyclylgroups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀),cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl(C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or contain a fused, bridged orspiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) andcan be saturated or can be partially unsaturated. “Carbocyclyl” alsoincludes ring systems wherein the carbocyclyl ring, as defined above, isfused with one or more aryl or heteroaryl groups wherein the point ofattachment is on the carbocyclyl ring, and in such instances, the numberof carbons continue to designate the number of carbons in thecarbocyclic ring system. In certain embodiments, each instance of acarbocyclyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 10 ring carbon atoms (“C₃₋₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). In certain embodiments,each instance of a cycloalkyl group is independently unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”)with one or more substituents. In certain embodiments, the cycloalkylgroup is unsubstituted C₃₋₁₀ cycloalkyl. In certain embodiments, thecycloalkyl group is substituted C₃₋₁₀ cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to10-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged or spiro ring system such as a bicyclic system(“bicyclic heterocyclyl”), and can be saturated or can be partiallyunsaturated. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. In certain embodiments, each instance of heterocyclyl isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-8 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl groupis a 5-6 membered non-aromatic ring system having ring carbon atoms and1-4 ring heteroatoms, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, and thiorenyl.Exemplary 4-membered heterocyclyl groups containing one heteroatominclude, without limitation, azetidinyl, oxetanyl, and thietanyl.Exemplary 5-membered heterocyclyl groups containing one heteroatominclude, without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl,and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, piperazinyl, morpholinyl,dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, triazinanyl.Exemplary 7-membered heterocyclyl groups containing one heteroatominclude, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₆ aryl ring (also referred toherein as a 5,6-bicyclic heterocyclic ring) include, without limitation,indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groupsfused to an aryl ring (also referred to herein as a 6,6-bicyclicheterocyclic ring) include, without limitation, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and the like.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclicor tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 πelectrons shared in a cyclic array) having 6-14 ring carbon atoms andzero heteroatoms provided in the aromatic ring system (“C₆₋₁₄ aryl”). Insome embodiments, an aryl group has six ring carbon atoms (“C₆ aryl”;e.g., phenyl). In some embodiments, an aryl group has ten ring carbonatoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). Insome embodiments, an aryl group has fourteen ring carbon atoms (“C₁₋₄aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein thearyl ring, as defined above, is fused with one or more carbocyclyl orheterocyclyl groups wherein the radical or point of attachment is on thearyl ring, and in such instances, the number of carbon atoms continue todesignate the number of carbon atoms in the aryl ring system. In certainembodiments, each instance of an aryl group is independently optionallysubstituted, e.g., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

“Heteroaryl” refers to a radical of a 5-10 membered monocyclic orbicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electronsshared in a cyclic array) having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, each instanceof a heteroaryl group is independently optionally substituted, e.g.,unsubstituted (“unsubstituted heteroaryl”) or substituted (“substitutedheteroaryl”) with one or more substituents. In certain embodiments, theheteroaryl group is unsubstituted 5-14 membered heteroaryl. In certainembodiments, the heteroaryl group is substituted 5-14 memberedheteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groupsinclude, without limitation, indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

“Partially unsaturated” refers to a group that includes at least onedouble or triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic groups (e.g., aryl or heteroaryl groups) asherein defined. Likewise, “saturated” refers to a group that does notcontain a double or triple bond, i.e., contains all single bonds.

In some embodiments, aliphatic, alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl groups, as defined herein, areoptionally substituted (e.g., “substituted” or “unsubstituted”aliphatic, “substituted” or “unsubstituted” alkyl, “substituted” or“unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl,“substituted” or “unsubstituted” carbocyclyl, “substituted” or“unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or“substituted” or “unsubstituted” heteroaryl group). In general, the term“substituted”, whether preceded by the term “optionally” or not, meansthat at least one hydrogen present on a group (e.g., a carbon ornitrogen atom) is replaced with a permissible substituent, e.g., asubstituent which upon substitution results in a stable compound, e.g.,a compound which does not spontaneously undergo transformation such asby rearrangement, cyclization, elimination, or other reaction. Unlessotherwise indicated, a “substituted” group has a substituent at one ormore substitutable positions of the group, and when more than oneposition in any given structure is substituted, the substituent iseither the same or different at each position. The term “substituted” iscontemplated to include substitution with all permissible substituentsof organic compounds, including any of the substituents described hereinthat results in the formation of a stable compound. The presentdisclosure contemplates any and all such combinations in order to arriveat a stable compound. For purposes of this disclosure, heteroatoms suchas nitrogen may have hydrogen substituents and/or any suitablesubstituent as described herein which satisfy the valencies of theheteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR^(aa), —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃,—OSi(R^(aa))₃—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa),—SC(═S)SR^(aa), —SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa),—SC(═O)R^(aa), —P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂,—OP(═O)(R^(aa))₂, —OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂,—OP(═O)₂N(R^(bb))₂, —P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂,—NR^(bb)P(═O)(OR^(cc))₂, —NR^(bb)P(═O)(NR^(bb))₂, —P(R^(cc))₂,—P(R^(cc))₃, —OP(R^(cc))₂, —OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂,—BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(aa) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups are joined to form a 3-14 memberedheterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R^(ff))₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,NR^(ff)SO₂R^(ee), —SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, 3-10membered heterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups, or two geminal R^(dd) substituents can be joined to form ═O or═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl,3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, ortwo R^(ff) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃—C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X⁻ is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions(e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate,benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate,naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonicacid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate,ethanoate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, and the like).

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—Cl), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quarternary nitrogenatoms. Exemplary nitrogen atom substitutents include, but are notlimited to, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R^(aa),—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include, but are not limited to, —OH,—OR^(aa), —N(R^(cc))₂, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa),—C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc),—SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl(e.g., aralkyl, heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aralkyl, aryl, and heteroaryl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb),R^(cc), and R^(dd) are as defined herein. Nitrogen protecting groups arewell known in the art and include those described in detail inProtecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts,3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include, but arenot limited to, formamide, acetamide, chloroacetamide,trichloroacetamide, trifluoroacetamide, phenylacetamide,3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide,3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) include, butare not limited to, methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) include,but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to,phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacylderivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanylderivative, N-acetylmethionine derivative,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N-(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to as a hydroxyl protectinggroup). Oxygen protecting groups include, but are not limited to,—R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃,—P(R^(cc))₂, —P(R^(cc))₃, —P(═O)₂R^(aa), —P(═O)(R^(aa))₂,—P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and —P(═O)(NR^(bb))₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Oxygen protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Exemplary oxygen protecting groups include, but are not limited to,methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate(TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec),2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutylcarbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts).

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a thiol protecting group).Sulfur protecting groups include, but are not limited to, —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃,—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and—P(═O)(NR^(bb))₂, wherein R^(aa), R^(bb), and R^(cc) are as definedherein. Sulfur protecting groups are well known in the art and includethose described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999,incorporated herein by reference.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The present disclosureis not intended to be limited in any manner by the above exemplarylisting of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al. describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (e.g., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or othernon-human animals, for example, non-human mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs), birds (e.g.,commercially relevant birds such as chickens, ducks, geese, and/orturkeys), rodents (e.g., rats and/or mice), reptiles, amphibians, andfish. In certain embodiments, the non-human animal is a mammal. Thenon-human animal may be a male or female at any stage of development. Anon-human animal may be a transgenic animal.

“Condition,” “disease,” and “disorder” are used interchangeably herein.

“Treat,” “treating” and “treatment” encompasses an action that occurswhile a subject is suffering from a condition which reduces the severityof the condition or retards or slows the progression of the condition(“therapeutic treatment”). “Treat,” “treating” and “treatment” alsoencompasses an action that occurs before a subject begins to suffer fromthe condition and which inhibits or reduces the severity of thecondition (“prophylactic treatment”).

An “effective amount” of a compound refers to an amount sufficient toelicit the desired biological response, e.g., treat the condition. Aswill be appreciated by those of ordinary skill in this art, theeffective amount of a compound described herein may vary depending onsuch factors as the desired biological endpoint, the pharmacokinetics ofthe compound, the condition being treated, the mode of administration,and the age and health of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound is an amountsufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “prophylactically effective amount” of a compound is an amountsufficient to prevent a condition, or one or more symptoms associatedwith the condition or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of a therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “methyltransferase” represents transferaseclass enzymes that are able to transfer a methyl group from a donormolecule to an acceptor molecule, e.g., an amino acid residue of aprotein or a nucleic base of a DNA molecule. Methylransferases typicallyuse a reactive methyl group bound to sulfur in S-adenosyl methionine(SAM) as the methyl donor. In some embodiments, a methyltransferasedescribed herein is a protein methyltransferase. In some embodiments, amethyltransferase described herein is a histone methyltransferase.Histone methyltransferases (HMT) are histone-modifying enzymes,(including histone-lysine N-methyltransferase and histone-arginineN-methyltransferase), that catalyze the transfer of one or more methylgroups to lysine and arginine residues of histone proteins. In certainembodiments, a methyltransferase described herein is a histone-arginineN-methyltransferase.

As generally described above, provided herein are compounds useful asPRMT5 inhibitors. In some embodiments, the present disclosure provides acompound of Formula (I):

or a pharmaceutically acceptable salt thereof,wherein

represents a single or double bond;

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

X is a bond, —O—, —N(R)—, —CR⁴R⁵—, —O—CR⁴R⁵, —N(R)—CR⁴R⁵—, —O—CR⁴R⁵—O—,—N(R)—CR⁴R⁵—O, —N(R)—CR⁴R⁵—N(R)—, —O—CR⁴R⁵—N(R)—, —CR⁴R⁵—O—,—CR⁴R⁵—N(R)—, —O—CR⁴R⁵—CR⁶R⁷—, —N(R)—CR⁴R⁵—CR⁶R⁷—, —CR⁶R⁷—CR⁴R⁵—O—,—CR⁶R⁷—CR⁴R⁵—N(R)—, or —CR⁶R⁷—CR⁴R⁵—;

each R is independently hydrogen or optionally substituted C₁₋₆aliphatic;

R² and R³ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R² andR³ are taken together with their intervening atoms to form an optionallysubstituted carbocyclic or heterocyclic ring;

R⁴ and R⁵ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁴ andR⁵ are taken together with their intervening atoms to form an optionallysubstituted carbocyclic or heterocyclic ring;

R⁶ and R⁷ are independently selected from the group consisting ofhydrogen, halo, —CN, —NO₂, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁶ andR⁷ are taken together with their intervening atoms to form an optionallysubstituted carbocyclic or heterocyclic ring;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

R⁸, R⁹, R¹⁰, and R¹¹ are independently hydrogen, halo, or optionallysubstituted aliphatic;

Cy is a monocyclic or bicyclic, saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy is substituted with 0, 1, 2, 3,or 4 R^(y) groups;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or an R^(y) group may be optionallytaken together with R² or R³ to form an optionally substituted 5- to6-membered carbocyclic or heterocyclic ring fused to Cy;

each R^(x) is independently selected from the group consisting of halo,—CN, optionally substituted aliphatic, —OR′, and —N(R″)₂;

R′ is hydrogen or optionally substituted aliphatic;

each R″ is independently hydrogen or optionally substituted aliphatic,or two R″ are taken together with their intervening atoms to form anoptionally substituted heterocyclic ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur; and

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits.

In certain embodiments, a provided compound is of Formula (I-a):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R⁸, R⁹, R¹⁰, R¹¹, R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (I-b):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R⁸, R⁹, R¹⁰, R¹¹, R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (I-c):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (I′):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (I′-a):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (I′-b):

or a pharmaceutically acceptable salt thereof, wherein R, R¹, R², R³,R^(x), n, X, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (II-a):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (II-b):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (III):

or a pharmaceutically acceptable salt thereof, wherein R, R², R³, R^(x),n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (III-a):

or a pharmaceutically acceptable salt thereof, wherein R, R², R³, R^(x),n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (III-b):

or a pharmaceutically acceptable salt thereof, wherein R, R², R³, R^(x),n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R⁴, R⁵,R^(x), n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (IV-a):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R⁴, R⁵,R^(x), n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (IV-b):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R⁴, R⁵,R^(x), n, and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (V-a):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In certain embodiments, a provided compound is of Formula (V-b):

or a pharmaceutically acceptable salt thereof, wherein R², R³, R^(x), n,and Cy are as described herein.

In some embodiments,

represents a single bond. In some embodiments,

represents a double bond.

As defined generally above, R¹ is hydrogen, R^(z), or —C(O) R^(z),wherein R^(z) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R¹ is hydrogen. In some embodiments, R¹ is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R¹ is methyl, ethyl, or propyl. In someembodiments, R¹ is —C(O) R^(z), wherein R^(z) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, R¹ is —C(O)R^(z), wherein R^(z) isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is acetyl.

As defined generally above, X is a bond, —O—, —N(R)—, —CR⁴R⁵—, —O—CR⁴R⁵,—N(R)—CR⁴R⁵—, —O—CR⁴R⁵—O—, —N(R)—CR⁴R⁵—O, —N(R)—CR⁴R⁵—N(R)—,—O—CR⁴R⁵—N(R)—, —CR⁴R⁵—O—, —CR⁴R⁵—N(R)—, —O—CR⁴R⁵—CR⁶R⁷—,—N(R)—CR⁴R⁵—CR⁶R⁷—, —CR⁶R⁷—CR⁴R⁵—O—, —CR⁶R⁷—CR⁴R⁵—N(R)—, or—CR⁶R⁷—CR⁴R⁵—. In certain embodiments, X is a bond, —O—, —N(R)—, or—CR⁴R⁵—, wherein R, R⁴, and R⁵ are as described herein. In certainembodiments, X is a bond. In certain embodiments, X is —O—. In someembodiments, X is —N(R)—. In certain embodiments, X is —NH—. In certainembodiments, X is —N(R)—, wherein R is optionally substituted C₁₋₆aliphatic. In certain embodiments, X is —N(R)—, wherein R is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, X is —N(R)—, wherein Ris unsubstituted C₁₋₆ alkyl. In certain embodiments, X is —N(Me)-. Insome embodiments, X is —CR⁴R⁵—. In certain embodiments, X is —CH₂—. Incertain embodiments, X is —CH₂—O—.

As defined generally above, each R is independently hydrogen oroptionally substituted C₁₋₆ aliphatic. In certain embodiments, R ishydrogen. In some embodiments, R is optionally substituted C₁₋₆aliphatic. In some embodiments, R is substituted C₁₋₆ aliphatic. In someembodiments, R is unsubstituted C₁₋₆ aliphatic. In some embodiments, Ris optionally substituted C₁₋₆ alkyl. In some embodiments, R issubstituted C₁₋₆ alkyl. In some embodiments, R is unsubstituted C₁₋₆alkyl. In some embodiments, R is methyl, ethyl, or propyl.

As defined generally above, R² and R³ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R² and R³ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring. In certain embodiments, R² and R³ are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A),—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R² and R³ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring.

In certain embodiments, R² is hydrogen. In some embodiments, R² is nothydrogen. In some embodiments, R² is halo. In certain embodiments, R² isfluoro. In some embodiments, R² is optionally substituted aliphatic. Incertain embodiments, R² is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R² is optionally substituted C₁₋₆ alkyl. In certainembodiments, R² is substituted C₁₋₆ alkyl. In certain embodiments, R² is—CF₃, CHF₂, or CH₂F. In certain embodiments, R² is unsubstituted C₁₋₆alkyl. In certain embodiments, R² is methyl, ethyl, or propyl. In someembodiments, R² is —CN or —NO₂. In some embodiments, R² is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R² is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R² is—N(R^(B))₂. In certain embodiments, R² is —NHR^(B). In certainembodiments, R² is —NH₂. In certain embodiments, R² is —OR^(A). Incertain embodiments, R² is —OH.

In certain embodiments, R³ is hydrogen. In some embodiments, R³ is nothydrogen. In some embodiments, R³ is halo. In certain embodiments, R³ isfluoro. In some embodiments, R³ is optionally substituted aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R³ is substituted C₁₋₆ alkyl. In certain embodiments, R³ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R³ is unsubstituted C₁₋₆alkyl. In certain embodiments, R³ is methyl, ethyl, or propyl. In someembodiments, R³ is —CN or —NO₂. In some embodiments, R³ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R³ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R³ is—N(R^(B))₂. In certain embodiments, R³ is —NHR^(B). In certainembodiments, R³ is —NH₂. In certain embodiments, R³ is —OR^(A). Incertain embodiments, R³ is —OH.

In some embodiments, R² and R³ are the same. In some embodiments, R² andR³ are different. In some embodiments, R² and R³ are each hydrogen. Insome embodiments, R² is hydrogen and R³ is not hydrogen. In someembodiments, R² is hydrogen and R³ is optionally substituted aliphatic.In some embodiments, R² is hydrogen and R³ is C₁₋₆ alkyl. In someembodiments, R² is hydrogen and R³ is methyl. In some embodiments, R² ishydrogen and R³ is ethyl or propyl. In some embodiments, R² is hydrogenand R³ is —CF₃, CHF₂, or CH₂F. In some embodiments, R² is hydrogen andR³ is —N(R^(B))₂ or —OR^(A). In some embodiments, R² is hydrogen and R³is —NH₂. In some embodiments, R² is hydrogen and R³ is —OH. In someembodiments, R² and R³ are not hydrogen. In some embodiments, R² and R³are independently optionally substituted aliphatic. In some embodiments,R² and R³ are methyl. In some embodiments, R² and R³ are taken togetherwith their intervening atoms to form an optionally substitutedcarbocyclic or heterocyclic ring.

As defined generally above, R⁴ and R⁵ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, OR^(A), N(R^(B))₂,) SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁴ and R⁵ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring. In certain embodiments, R⁴ and R⁵ are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), N(R^(B))₂, SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A),—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁴ and R⁵ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring.

In certain embodiments, R⁴ is hydrogen. In some embodiments, R⁴ is nothydrogen. In some embodiments, R⁴ is halo. In certain embodiments, R⁴ isfluoro. In some embodiments, R⁴ is optionally substituted aliphatic. Incertain embodiments, R⁴ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R⁴ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁴ is substituted C₁₋₆ alkyl. In certain embodiments, R⁴ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R⁴ is unsubstituted C₁₋₆alkyl. In certain embodiments, R⁴ is methyl, ethyl, or propyl. In someembodiments, R⁴ is —CN or —NO₂. In some embodiments, R⁴ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R⁴ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, NR^(B)C(═S)R^(A), S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁴ is—N(R^(B))₂. In certain embodiments, R⁴ is —NHR^(B). In certainembodiments, R⁴ is —NH₂. In certain embodiments, R⁴ is —OR^(A). Incertain embodiments, R⁴ is —OH.

In certain embodiments, R⁵ is hydrogen. In some embodiments, R⁵ is nothydrogen. In some embodiments, R⁵ is halo. In certain embodiments, R⁵ isfluoro. In some embodiments,

R⁵ is optionally substituted aliphatic. In certain embodiments, R⁵ isoptionally substituted C₁₋₆ aliphatic. In certain embodiments, R⁵ isoptionally substituted C₁₋₆ alkyl. In certain embodiments, R⁵ issubstituted C₁₋₆ alkyl. In certain embodiments, R⁵ is —CF₃, CHF₂, orCH₂F. In certain embodiments, R⁵ is unsubstituted C₁₋₆ alkyl. In certainembodiments, R⁵ is methyl, ethyl, or propyl. In some embodiments, R⁵ is—CN or —NO₂. In some embodiments, R⁵ is optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, or optionally substituted heteroaryl. In some embodiments,R⁵ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁵ is—N(R^(B))₂. In certain embodiments, R⁵ is —NHR^(B). In certainembodiments, R⁵ is —NH₂. In certain embodiments, R⁵ is —OR^(A). Incertain embodiments, R⁵ is —OH.

In some embodiments, R⁴ and R⁵ are the same. In some embodiments, R⁴ andR⁵ are different. In some embodiments, R⁴ and R⁵ are each hydrogen. Insome embodiments, R⁴ is hydrogen and R⁵ is not hydrogen. In someembodiments, R⁴ is hydrogen and R⁵ is optionally substituted aliphatic.In some embodiments, R⁴ is hydrogen and R⁵ is C₁₋₆ alkyl. In someembodiments, R⁴ is hydrogen and R⁵ is methyl. In some embodiments, R⁴ ishydrogen and R⁵ is ethyl or propyl. In certain embodiments, R⁴ andhydrogen and R⁵ is —CF₃, CHF₂, or CH₂F. In some embodiments, R⁴ ishydrogen and R⁵ is —N(R^(B))₂ or —OR^(A). In some embodiments, R⁴ ishydrogen and R⁵ is —NH₂. In some embodiments, R⁴ is hydrogen and R⁵ is—OH. In some embodiments, R⁴ and R⁵ are not hydrogen. In someembodiments, R⁴ and R⁵ are independently optionally substitutedaliphatic. In some embodiments, R⁴ and R⁵ are methyl. In someembodiments, R⁴ and R⁵ are taken together with their intervening atomsto form an optionally substituted carbocyclic or heterocyclic ring.

As defined generally above, R⁶ and R⁷ are independently selected fromthe group consisting of hydrogen, halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted phenyl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁶ and R⁷ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring. In certain embodiments, R⁶ and R⁷ are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A),—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁶ and R⁷ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring.

In certain embodiments, R⁶ is hydrogen. In some embodiments, R⁶ is nothydrogen. In some embodiments, R⁶ is halo. In certain embodiments, R⁶ isfluoro. In some embodiments, R⁶ is optionally substituted aliphatic. Incertain embodiments, R⁶ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R⁶ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁶ is substituted C₁₋₆ alkyl. In certain embodiments, R⁶ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R⁶ is unsubstituted C₁₋₆alkyl. In certain embodiments, R⁶ is methyl, ethyl, or propyl. In someembodiments, R⁶ is —CN or —NO₂. In some embodiments, R⁶ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R⁶ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁶ is—N(R^(B))₂. In certain embodiments, R⁶ is —NHR^(B). In certainembodiments, R⁶ is —NH₂. In certain embodiments, R⁶ is —OR^(A). Incertain embodiments, R⁶ is —OH.

In certain embodiments, R⁷ is hydrogen. In some embodiments, R⁷ is nothydrogen. In some embodiments, R⁷ is halo. In certain embodiments, R⁷ isfluoro. In some embodiments, R⁷ is optionally substituted aliphatic. Incertain embodiments, R⁷ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R⁷ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁷ is substituted C₁₋₆ alkyl. In certain embodiments, R⁷ is—CF₃, CHF₂, or CH₂F. In certain embodiments, R⁷ is unsubstituted C₁₋₆alkyl. In certain embodiments, R⁷ is methyl, ethyl, or propyl. In someembodiments, R⁷ is —CN or —NO₂. In some embodiments, R⁷ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R⁷ is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In certain embodiments, R⁷ is—N(R^(B))₂. In certain embodiments, R⁷ is —NHR^(B). In certainembodiments, R⁷ is —NH₂. In certain embodiments, R⁷ is —OR^(A). Incertain embodiments, R⁷ is —OH.

In some embodiments, R⁶ and R⁷ are the same. In some embodiments, R⁶ andR⁷ are different. In some embodiments, R⁶ and R⁷ are each hydrogen. Insome embodiments, R⁶ is hydrogen and R⁷ is not hydrogen. In someembodiments, R⁶ is hydrogen and R⁷ is optionally substituted aliphatic.In some embodiments, R⁶ is hydrogen and R⁷ is C₁₋₆ alkyl. In someembodiments, R⁶ is hydrogen and R⁷ is methyl. In some embodiments, R⁶ ishydrogen and R⁷ is ethyl or propyl. In certain embodiments, R⁶ andhydrogen and R⁷ is —CF₃, CHF₂, or CH₂F. In some embodiments, R⁶ ishydrogen and R⁷ is —N(R^(B))₂ or —OR^(A). In some embodiments, R⁶ ishydrogen and R⁷ is —NH₂. In some embodiments, R⁶ is hydrogen and R⁷ is—OH. In some embodiments, R⁶ and R⁷ are not hydrogen. In someembodiments, R⁶ and R⁷ are independently optionally substitutedaliphatic. In some embodiments, R⁶ and R⁷ are methyl. In someembodiments, R⁶ and R⁷ are taken together with their intervening atomsto form an optionally substituted carbocyclic or heterocyclic ring.

As defined generally above, R⁸, R⁹, R¹⁰, and R¹¹ are independentlyhydrogen, halo, or optionally substituted aliphatic. In someembodiments, R⁸, R⁹, R¹⁰, and R¹¹ are hydrogen. In some embodiments, R⁹,R¹⁰, and R¹¹ are hydrogen, and R⁸ is optionally substituted aliphatic.In some embodiments, R⁹, R¹⁰, and R¹¹ are hydrogen, and R⁸ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁹, R¹⁰, and R¹¹ arehydrogen, and R⁸ is optionally substituted C₁₋₃ aliphatic. In someembodiments, R⁹, R¹⁰, and R¹¹ are hydrogen, and R⁸ is methyl. In someembodiments, R⁸, R⁹, and R¹⁰ are hydrogen, and R¹¹ is optionallysubstituted aliphatic. In some embodiments, R⁸, R⁹, and R¹⁰ arehydrogen, and R¹¹ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R⁸, R⁹, and R¹⁰ are hydrogen, and R¹¹ is optionallysubstituted C₁₋₃ aliphatic. In some embodiments, R⁸, R⁹, and R¹⁰ arehydrogen, and R¹¹ is methyl. In some embodiments, R⁸ is hydrogen. Insome embodiments, R⁸ is halo. In certain embodiments, R⁸ is fluoro. Insome embodiments, R⁸ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R⁸ is optionally substituted C₁₋₃ alkyl. In certainembodiments, R⁸ is methyl. In some embodiments, R⁹ is hydrogen. In someembodiments, R⁹ is halo. In certain embodiments, R⁹ is fluoro. In someembodiments, R⁹ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R⁹ is optionally substituted C₁₋₃ alkyl. In certainembodiments, R⁹ is methyl. In some embodiments, R¹⁰ is hydrogen. In someembodiments, R¹⁰ is halo. In certain embodiments, R¹⁰ is fluoro. In someembodiments, R¹⁰ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R¹⁰ is optionally substituted C₁₋₃ alkyl. In certainembodiments, R¹⁰ is methyl. In some embodiments, R¹¹ is hydrogen. Insome embodiments, R¹¹ is halo. In certain embodiments, R¹¹ is fluoro. Insome embodiments, R¹¹ is optionally substituted C₁₋₆ aliphatic. In someembodiments, R¹¹ is optionally substituted C₁₋₃ alkyl. In certainembodiments, R¹¹ is methyl.

As defined generally above, Cy is a monocyclic or bicyclic, saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein Cy issubstituted with 0, 1, 2, 3, or 4 R^(y) groups. In certain embodiments,Cy is unsubstituted. In certain embodiments, Cy is substituted with oneor two R^(y) groups. In certain embodiments, Cy is substituted with oneR^(y) group. In certain embodiments, Cy is substituted with two R^(y)groups. In certain embodiments, Cy is substituted with three R^(y)groups. In certain embodiments, Cy is substituted with four R^(y)groups.

In certain embodiments, Cy is phenyl substituted with 0, 1, 2, 3, or 4R^(y) groups. In certain embodiments, Cy is phenyl substituted with oneor two R^(y) groups. In certain embodiments, Cy is unsubstituted phenyl.In certain embodiments, Cy is phenyl substituted with one R^(y) group.In certain embodiments, Cy is phenyl substituted with two R^(y) groups.In certain embodiments, Cy is phenyl substituted with three R^(y)groups. In certain embodiments, Cy is phenyl substituted with four R^(y)groups.

In certain embodiments, Cy is a 5- to 6-membered heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur,and is substituted with 0, 1, 2, 3, or 4 R^(y) groups. In certainembodiments, Cy is an unsubstituted 5- to 6-membered heteroaryl having1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Cy is a 5- to 6-membered heteroarylhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, andsulfur, and is substituted with one or two R^(y) groups. In certainembodiments, Cy is a 5- to 6-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, and issubstituted with one R^(y) group. In certain embodiments, Cy is a5-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur (e.g., furanyl, thienyl, pyrrolyl,oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyrazolyl, isothiazolyl,triazolyl, oxadiazolyl, thiadiazolyl), and is substituted with 0, 1, 2,3, or 4 R^(y) groups. In certain embodiments, Cy is a 6-memberedheteroaryl having 1-3 nitrogens (e.g., pyridyl, pyrimidyl, pyridazinyl,pyrazinyl, triazinyl), and is substituted with 0, 1, 2, 3, or 4 R^(y)groups.

In certain embodiments, Cy is a bicyclic saturated, partiallyunsaturated, or aromatic ring having 0-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, wherein Cy is substitutedwith 0, 1, 2, 3, or 4 R^(y) groups. In certain embodiments, Cy is an 8-to 12-membered bicyclic saturated, partially unsaturated, or aromaticring having 0-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4R^(y) groups. In certain embodiments, Cy is an unsubstituted bicyclicsaturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, Cy is a bicyclic saturated, partially unsaturated,or aromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy is substituted with one or twoR^(y) groups. In certain embodiments, Cy is a bicyclic saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur, wherein Cy issubstituted with one R^(y) group. In certain embodiments, Cy is abicyclic saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Cy is substituted with two R^(y) groups. In certain embodiments,Cy is a bicyclic saturated, partially unsaturated, or aromatic ringhaving 0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur,

wherein Cy is substituted with three R^(y) groups. In certainembodiments, Cy is a bicyclic saturated, partially unsaturated, oraromatic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, wherein Cy is substituted with four R^(y)groups.

In certain embodiments, Cy is an 8- to 10-membered bicyclic heteroarylhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R^(y) groups. Incertain embodiments, Cy is a 9-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur(e.g., indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl,isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl,benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl,benzisothiazolyl, benzthiadiazolyl, indolizinyl), wherein Cy issubstituted with 0, 1, 2, 3, or 4 R^(y) groups. In certain embodiments,Cy is a 10-membered bicyclic heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur (e.g.,naphthyridinyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl),wherein Cy is substituted with 0, 1, 2, 3, or 4 R^(y) groups. In certainembodiments, Cy is selected from the group consisting of quinoline,benzimidazole, benzopyrazole, quinoxaline, tetrahydroquinoline,tetrahydroisoquinoline, naphthalene, tetrahydronaphthalene,2,3-dihydrobenzo[b][1,4]dioxine, isoindole,2H-benzo[b][1,4]oxazin-3(4H)-one, 3,4-dihydro-2H-benzo[b][1,4]oxazine,and quinoxalin-2(1H)-one, wherein Cy is substituted with 0, 1, 2, 3, or4 R^(y) groups.

In certain embodiments, Cy is a 5,6-fused bicyclic heteroaryl ringsystem such as one of the following:

In any bicyclic heteroaryl group shown above, the point of attachmentcan be any carbon or nitrogen atom, as valency permits, and the ring maybe substituted with 0, 1, 2, 3, 4, or 5 R^(y) groups, as valencypermits.

In certain embodiments, a provided compound is of Formula (VII):

or a pharmaceutically acceptable salt thereof, wherein Cy is a 5,6-fusedbicyclic heteroaryl as described herein, wherein Cy is substituted with0, 1, 2, 3, 4, or 5 R^(y) groups, as valency permits.

In certain embodiments, a provided compound is of Formula (VII-a):

or a pharmaceutically acceptable salt thereof, wherein Cy is a 5,6-fusedbicyclic heteroaryl as described herein, wherein Cy is substituted with0, 1, 2, 3, 4, or 5 R^(y) groups, as valency permits.

In certain embodiments, a provided compound is of Formula (VII-b):

or a pharmaceutically acceptable salt thereof, wherein Cy is a 5,6-fusedbicyclic heteroaryl as described herein, wherein Cy is substituted with0, 1, 2, 3, 4, or 5 R^(y) groups, as valency permits.

As defined generally above, each R^(y) is independently selected fromthe group consisting of halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(RB)₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein; or an R^(y) group may be optionally taken togetherwith R² or R³ to form an optionally substituted 5- to 6-memberedcarbocyclic or heterocyclic ring fused to Cy. In some embodiments, eachR^(y) is independently selected from the group consisting of halo, —CN,—NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted phenyl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—OC(O)R^(A), —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein.

In some embodiments, at least one R^(y) is halo. In certain embodiments,at least one R^(y) is fluoro. In certain embodiments, at least one R^(y)is chloro. In some embodiments, at least one R^(y) is —CN. In someembodiments, at least one R^(y) is —OR^(A), wherein R^(A) is optionallysubstituted aliphatic. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, at least one R^(y) is methoxy, ethoxy, or propoxy. Incertain embodiments, at least one R^(y) is methoxy. In some embodiments,at least one R^(y) is —OR^(A), wherein R^(A) is substituted C₁₋₆ alkyl.In certain embodiments, at least one R^(y) is —OCH₂CH₂N(CH₃)₂. In someembodiments, at least one R^(y) is —N(R^(B))₂. In some embodiments, atleast one R^(y) is —N(R^(B))₂, wherein each R^(B) is independentlyselected from hydrogen or C₁₋₆ alkyl. In some embodiments, at least oneR^(y) is —NHR^(B). In some embodiments, at least one R^(y) is —N(C₁₋₆alkyl)₂, —NH(C₁₋₆ alkyl), or —NH₂. In certain embodiments, at least oneR^(y) is —NH₂. In certain embodiments, at least one R^(y) is —NHCH₃. Incertain embodiments, at least one R^(y) is —N(CH₃)₂.

In some embodiments, at least one R^(y) is optionally substitutedaliphatic. In certain embodiments, at least one R^(y) is substitutedaliphatic. In certain embodiments, at least one R^(y) is unsubstitutedaliphatic. In some embodiments, at least one R^(y) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) isunsubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) issubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) ismethyl, ethyl, or propyl. In certain embodiments, at least one R^(y) ismethyl. In certain embodiments, at least one R^(y) is —CF₃, CHF₂, orCH₂F. In certain embodiments, at least one R^(y) is C₁₋₆ alkylsubstituted with aryl, heteroaryl, or heterocyclyl. In certainembodiments, at least one R^(y) is benzyl. In certain embodiments, atleast one R^(y) is —(C₁₋₆ alkyl)-heteroaryl. In certain embodiments, atleast one R^(y) is —(C₁₋₆ alkyl)-heterocyclyl. In certain embodiments,at least one R^(y) is —CH₂-heteroaryl. In certain embodiments, at leastone R^(y) is —CH₂-heterocyclyl.

In some embodiments, at least one R^(y) is —C(O)N(R^(B))₂. In certainembodiments, at least one R^(y) is —C(O)NHR^(B). In certain embodiments,at least one R^(y) is —C(O)NH₂. In certain embodiments, at least oneR^(y) is —C(O)N(R^(B))₂, wherein the R^(B) groups are taken togetherwith their intervening atoms to form an optionally substituted 5- to6-membered heterocyclyl. In certain embodiments, at least one R^(y) is—C(O)N(R^(B))₂, wherein the R^(B) groups are taken together with theirintervening atoms to form an optionally substituted morpholinyl.

In some embodiments, at least one R^(y) is —SO₂N(R^(B))₂. In certainembodiments, at least one R^(y) is —SO₂NHR^(B). In certain embodiments,at least one R^(y) is —SO₂NH₂. In certain embodiments, at least oneR^(y) is —SO₂N(R^(B))₂, wherein neither R^(B) is hydrogen. In certainembodiments, at least one R^(y) is —SO₂NH(C₁₋₆ alkyl) or —SO₂N(C₁₋₆alkyl)₂. In certain embodiments, at least one R^(y) is —SO₂N(CH₃)₂. Incertain embodiments, at least one R^(y) is —SO₂N(R^(B))₂, wherein theR^(B) groups are taken together with their intervening atoms to form anoptionally substituted 5- to 6-membered heterocyclyl. In certainembodiments, at least one R^(y) is —SO₂-morpholinyl. In certainembodiments, at least one R^(y) is —SO₂-piperidinyl, —SO₂-piperazinyl,or —SO₂-piperidinyl.

In some embodiments, at least one R^(y) is —SO₂R^(A). In someembodiments, at least one R^(y) is —SO₂R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —SO₂(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—SO₂CH₃. In some embodiments, at least one R^(y) is —C(O)R^(A). In someembodiments, at least one R^(y) is —C(O)R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —C(O)(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—C(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))C(O)R^(A). Incertain embodiments, at least one R^(y) is —NHC(O)R^(A). In certainembodiments, at least one R^(y) is —NHC(O)(C₁₋₆ alkyl). In certainembodiments, at least one R^(y) is —NHC(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))SO₂R^(A). In someembodiments, at least one R^(y) is —NHSO₂R^(A). In some embodiments, atleast one R^(y) is —N(C₁₋₆ alkyl)SO₂R^(A). In certain embodiments, atleast one R^(y) is —NHSO₂(C₁₋₆ alkyl) or —N(C₁₋₆ alkyl)SO₂(C₁₋₆ alkyl).In certain embodiments, at least one R^(y) is —NHSO₂CH₃. In certainembodiments, at least one R^(y) is —N(CH₃)SO₂CH₃.

In some embodiments, at least one R^(y) is optionally substitutedheterocyclyl, optionally substituted carbocyclyl, optionally substitutedaryl, or optionally substituted heteroaryl. In certain embodiments, atleast one R^(y) is an optionally substituted 5- to 6-memberedheterocyclyl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heterocyclyl having oneheteroatom selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is optionally substituted pyrrolidinyl.In certain embodiments, at least one R^(y) is pyrroldinyl,hydroxypyrrolidinyl, or methylpyrrolidinyl. In certain embodiments, atleast one R^(y) is an optionally substituted 6-membered heterocyclylhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is an optionallysubstituted 6-membered heterocyclyl having one heteroatom selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is optionally substituted piperidinyl. In certain embodiments, at leastone R^(y) is an optionally substituted 6-membered heterocyclyl havingtwo heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is optionallysubstituted piperidinyl, optionally substituted piperazinyl, oroptionally substituted morpholinyl. In certain embodiments, at least oneR^(y) is morpholinyl, tetrahydropyranyl, piperidinyl, methylpiperidinyl,piperazinyl, methylpiperazinyl, acetylpiperazinyl,methylsulfonylpiperazinyl, aziridinyl, or methylaziridinyl. In someembodiments, at least one R^(y) is an optionally substituted 5- to6-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, at least one R^(y) is an optionally substituted5-membered heteroaryl having one heteroatom selected from nitrogen,oxygen, and sulfur. In certain embodiments, at least one R^(y) is anoptionally substituted 5-membered heteroaryl having two heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is an optionally substituted 6-memberedheteroaryl having 1-3 nitrogens. In certain embodiments, at least oneR^(y) is an optionally substituted pyrazolyl. In certain embodiments, atleast one R^(y) is an optionally substituted imidazolyl. In certainembodiments, at least one R^(y) is an optionally substituted pyridyl. Incertain embodiments, at least one R^(y) is an optionally substitutedpyrimidyl. In certain embodiments, at least one R^(y) is pyrazolyl,methylpyrazolyl, imidazolyl, or methylimidazolyl.

In some embodiments, an R^(y) group is taken together with R² or R³ andtheir intervening atoms to form a 5- to 6-membered carbocyclic orheterocyclic ring fused to Cy.

In certain embodiments, Cy is selected from the group consisting of:

In some embodiments, Cy is selected from the group consisting of:

In some embodiments, Cy is selected from the group consisting of:

As defined generally above, each R^(x) is independently selected fromthe group consisting of halo, —CN, optionally substituted aliphatic,—OR′, and —N(R″)₂. In certain embodiments, at least one R^(x) is halo.In certain embodiments, at least one R^(x) is fluoro. In certainembodiments, at least one R^(x) is —CN. In certain embodiments, at leastone R^(x) is optionally substituted aliphatic. In certain embodiments,at least one R^(x) is optionally substituted C₁₋₆ alkyl. In certainembodiments, at least one R^(x) is methyl. In certain embodiments, atleast one R^(x) is —CF₃. In certain embodiments, at least one R^(x) is—OR′ or —N(R″)₂. In certain embodiments, R^(x) is not —OR′ or —N(R″)₂.In certain embodiments, at least one R^(x) is —OCH₃. In certainembodiments, R^(x) is not —OCH₃.

One of ordinary skill in the art will appreciate that an R^(x) group canbe attached anywhere on the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the benzene portion of the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the tetrahydropyridine or dihydropyridine portion of thetetrahydroisoquinoline or dihydroisoquinoline ring. In certainembodiments, R^(x) groups are attached to both the benzene portion andthe tetrahydropyridine (or dihydropyridine) portion of thetetrahydroisoquinoline (or dihydroisoquinoline) ring. See, for example,the structures shown below:

In certain embodiments, a provided compound is of Formula (VI):

or a pharmaceutically acceptable salt thereof.

As defined generally above, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. Incertain embodiments, n is 0. In certain embodiments, n is 1. In certainembodiments, n is 2.

In certain embodiments, a provided compound is a compound listed inTable 1, or a pharmaceutically acceptable salt thereof.

TABLE 1 Exemplary Compounds LC-MS Cmpd m/z No Structure Exact mass (M +H) 1

340.1787 341.2 2

338.1994 339.2 3

352.2151 353.2 4

485.2678 486.2 5

354.1943 355.1 6

391.1896 392.2 7

358.1693 359.1 8

365.1739 366.1 9

354.1943 355.2 10

358.1693 359.2 11

365.1739 366.2 12

370.1893 371.2 13

383.1845 384.2 14

418.1562 419.2 15

354.1943 355.1 16

358.1693 359.1 17

365.1739 366.1 18

339.1947 340.1 19

354.1943 355.2 20

370.1893 371.1 21

397.2002 398.2 22

370.1893 371.1 23

394.2005 395.1 24

346.2256 347.2 25

368.21 369.2 26

354.1943 355.2 27

383.2209 384.2 28

433.1671 434.1 29

383.1845 384.2 30

394.2005 395.1 31

397.2002 398.1 32

340.1787 341.2 33

340.1787 341.2 34

383.1845 384.1 35

383.2209 384.2 36

397.2002 398.1 37

418.1562 419.1 38

383.2209 384.2 39

405.2052 406.2 40

422.2569 423.2 41

348.2049 349.2 42

433.1671 434.1 43

419.2209 420.2 44

391.1896 392.1 45

391.1896 392.1 46

394.2005 395.2 47

418.1562 419.1 48

476.2424 477.2 49

425.2315 426.2 50

406.2005 407.2 51

325.179 326.1 52

419.1515 420.1 53

433.1671 434.1 54

404.2212 405.2 55

325.1838 325.1 56

330.2307 331.2 57

324.1838 325.1 58

330.2307 331.2 59

339.1947 340.2 60

375.1947 376.1 61

339.1947 340.1 62

339.1947 340.1 63

339.1947 340.1 64

394.2256 395.1 65

419.1515 420.1 66

390.1943 391.2 67

406.2005 407.2 68

406.2005 407.2 69

393.2165 394.2 70

463.2583 464.2 71

477.274 478.3 72

405.2052 406.2 73

405.2052 406.1 74

405.2052 406.1 75

439.2471 440.2 76

405.2052 406.2 77

425.2315 426.2 78

425.2315 426.2 79

424.2474 425.2 80

425.1951 426.2 81

411.2158 412.1 82

394.2005 395.2 83

453.2264 454.2 84

438.2631 439.3 85

438.2631 439.3 86

469.2577 470.2 87

469.2577 470.2 88

427.2471 428.1 89

427.2471 428.2 90

466.258 467.2 91

395.2209 396.1 92

392.1848 393.2 93

439.2471 440.2 94

490.258 491.2 95

449.2427 450.2 96

463.2583 464.2 97

421.2002 422.2 98

421.2002 422.2 99

409.2365 410.2 100

398.1842 399.1 101

437.2315 438.2 102

473.1984 474.2 103

423.2522 424.3 104

434.2318 435.2 105

434.2318 435.2 106

502.225 503.2 107

488.2093 489.2 108

502.225 503.2 109

447.1828 448.1 110

473.1984 474.1 111

489.1934 490.1 112

397.2002 398.2 113

411.2158 412.2 114

432.1719 433.1 115

406.2005 407.1 116

409.2365 410.2 117

423.2522 424.2 118

394.2005 395.2 119

395.2209 396.1 120

409.2365 410.2 121

440.2424 441.2 122

530.2199 531.2 123

433.1671 434.1 124

411.1794 412.2 125

408.1798 409.2 126

433.1671 434.1 127

384.2161 385.1 128

398.1954 399.1 129

408.1798 409.1 130

437.2678 438.3 131

419.1515 420.1 132

395.2209 396.2 133

384.2161 385.2 134

437.2678 438.3 135

409.2365 410.3 136

423.2522 424.2 137

406.2005 407.1 138

420.2161 421.1 139

434.1624 435.1 140

434.1624 435.1 141

455.242 456.2 142

447.1828 448.2 143

384.2161 385.2 144

427.1707 428.2 145

356.1848 357.1 146

407.2209 408.2 147

398.1954 399.2 148

398.1954 399.2 149

489.1934 490.1 150

395.2209 396.2 151

409.2365 410.2 152

355.1896 356.2 153

420.2161 421.2 154

392.1848 393.2 155

447.1828 448.1 156

420.2161 421.2 157

447.1828 448.1 158

408.2161 409.2 159

394.2005 395.2 160

394.2005 395.1 161

454.2216 455.2 162

398.1954 399.2 163

407.2209 408.2 164

407.2209 408.2 165

340.1787 341.1 166

348.2049 349.1 167

377.2315 378.2 168

407.1957 408.2 169

462.1937 463.1 170

433.1671 434.1 171

334.1893 335.1 172

440.2424 441.2 173

354.1943 355.1 174

368.1736 369.2 175

378.2056 379.1 176

390.2631 391.1 177

351.1947 352.1 178

364.2151 365.1 179

364.2151 365.1 180

364.2151 365.1 181

392.1848 393.2 182

363.1947 364.1 183

368.1736 369.2 184

357.2416 358.1 185

365.2103 366.1 186

406.2005 407.1 187

420.2161 421.1 188

496.2686 497.3 189

477.2376 478.3 190

438.2631 439.3 191

504.2406 505.1 192

378.2056 379.1 193

395.1667 396.2

In certain embodiments, a provided compound inhibits PRMT5. In certainembodiments, a provided compound inhibits wild-type PRMT5. In certainembodiments, a provided compound inhibits a mutant PRMT5. In certainembodiments, a provided compound inhibits PRMT5, e.g., as measured in anassay described herein. In certain embodiments, the PRMT5 is from ahuman. In certain embodiments, a provided compound inhibits PRMT5 at anIC₅₀ less than or equal to 10 μM. In certain embodiments, a providedcompound inhibits PRMT5 at an IC₅₀ less than or equal to 1 μM. Incertain embodiments, a provided compound inhibits PRMT5 at an IC₅₀ lessthan or equal to 0.1 μM. In certain embodiments, a provided compoundinhibits PRMT5 in a cell at an EC₅₀ less than or equal to 10 μM. Incertain embodiments, a provided compound inhibits PRMT5 in a cell at anEC₅₀ less than or equal to 1 μM. In certain embodiments, a providedcompound inhibits PRMT5 in a cell at an EC₅₀ less than or equal to 0.1μM. In certain embodiments, a provided compound inhibits cellproliferation at an EC₅₀ less than or equal to 10 μM. In certainembodiments, a provided compound inhibits cell proliferation at an EC₅₀less than or equal to 1 μM. In certain embodiments, a provided compoundinhibits cell proliferation at an EC₅₀ less than or equal to 0.1 μM. Insome embodiments, a provided compound is selective for PRMT5 over othermethyltransferases. In certain embodiments, a provided compound is atleast about 10-fold selective, at least about 20-fold selective, atleast about 30-fold selective, at least about 40-fold selective, atleast about 50-fold selective, at least about 60-fold selective, atleast about 70-fold selective, at least about 80-fold selective, atleast about 90-fold selective, or at least about 100-fold selective forPRMT5 relative to one or more other methyltransferases.

It will be understood by one of ordinary skill in the art that the PRMT5can be wild-type PRMT5, or any mutant or variant of PRMT5.

In certain embodiments, the PRMT5 is isoform A (GenBank accession no.NP006100) (SEQ ID NO.: 1):

MAAMAVGGAG GSRVSSGRDL NCVPEIADTL GAVAKQGFDFLCMPVFHPRF KREFIQEPAK NRPGPQTRSD LLLSGRDWNTLIVGKLSPWI RPDSKVEKIR RNSEAAMLQE LNFGAYLGLPAFLLPLNQED NTNLARVLTN HIHTGHHSSM FWMRVPLVAPEDLRDDIIEN APTTHTEEYS GEEKTWMWWH NFRTLCDYSKRIAVALEIGA DLPSNHVIDR WLGEPIKAAI LPTSIFLTNKKGFPVLSKMH QRLIFRLLKL EVQFIITGTN HHSEKEFCSYLQYLEYLSQN RPPPNAYELF AKGYEDYLQS PLQPLMDNLESQTYEVFEKD PIKYSQYQQA IYKCLLDRVP EEEKDTNVQVLMVLGAGRGP LVNASLRAAK QADRRIKLYA VEKNPNAVVTLENWQFEEWG SQVTVVSSDM REWVAPEKAD IIVSELLGSFADNELSPECL DGAQHFLKDD GVSIPGEYTS FLAPISSSKLYNEVRACREK DRDPEAQFEM PYVVRLHNFH QLSAPQPCFTFSHPNRDPMI DNNRYCTLEF PVEVNTVLHG FAGYFETVLYQDITLSIRPE THSPGMFSWF PILFPIKQPI TVREGQTICVRFWRCSNSKK VWYEWAVTAP VCSAIHNPTG RSYTIGL

In certain embodiments, the PRMT5 is isoform B (GenBank accession no.NP001034708) (SEQ ID NO.: 2)

MRGPNSGTEK GRLVIPEKQG FDFLCMPVFH PRFKREFIQEPAKNRPGPQT RSDLLLSGRD WNTLIVGKLS PWIRPDSKVEKIRRNSEAAM LQELNFGAYL GLPAFLLPLN QEDNTNLARVLTNHIHTGHH SSMFWMRVPL VAPEDLRDDI IENAPTTHTEEYSGEEKTWM WWHNFRTLCD YSKRIAVALE IGADLPSNHVIDRWLGEPIK AAILPTSIFL TNKKGFPVLS KMHQRLIFRLLKLEVQFIIT GTNHHSEKEF CSYLQYLEYL SQNRPPPNAYELFAKGYEDY LQSPLQPLMD NLESQTYEVF EKDPIKYSQYQQAIYKCLLD RVPEEEKDTN VQVLMVLGAG RGPLVNASLRAAKQADRRIK LYAVEKNPNA VVTLENWQFE EWGSQVTVVSSDMREWVAPE KADIIVSELL GSFADNELSP ECLDGAQHFLKDDGVSIPGE YTSFLAPISS SKLYNEVRAC REKDRDPEAQFEMPYVVRLH NFHQLSAPQP CFTFSHPNRD PMIDNNRYCTLEFPVEVNTV LHGFAGYFET VLYQDITLSI RPETHSPGMFSWFPILFPIK QPITVREGQT ICVRFWRCSN SKKVWYEWAV TAPVCSAIHN PTGRSYTIGL

In certain embodiments, the PRMT5 is transcript variant 1 (GenBankaccession no. NM_(—)006109).

The present disclosure provides pharmaceutical compositions comprising acompound described herein, e.g., a compound of Formula (I), or apharmaceutically acceptable salt thereof, as described herein, andoptionally a pharmaceutically acceptable excipient. It will beunderstood by one of ordinary skill in the art that the compoundsdescribed herein, or salts thereof, may be present in various forms,such as amorphous, hydrates, solvates, or polymorphs. In certainembodiments, a provided composition comprises two or more compoundsdescribed herein. In certain embodiments, a compound described herein,or a pharmaceutically acceptable salt thereof, is provided in aneffective amount in the pharmaceutical composition. In certainembodiments, the effective amount is a therapeutically effective amount.In certain embodiments, the effective amount is an amount effective forinhibiting PRMT5. In certain embodiments, the effective amount is anamount effective for treating a PRMT5-mediated disorder. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the effective amount is an amounteffective to prevent a PRMT5-mediated disorder.

Pharmaceutically acceptable excipients include any and all solvents,diluents, or other liquid vehicles, dispersions, suspension aids,surface active agents, isotonic agents, thickening or emulsifyingagents, preservatives, solid binders, lubricants, and the like, assuited to the particular dosage form desired. General considerations informulation and/or manufacture of pharmaceutical compositions agents canbe found, for example, in Remington's Pharmaceutical Sciences, SixteenthEdition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), andRemington: The Science and Practice of Pharmacy, 21st Edition(Lippincott Williams & Wilkins, 2005).

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing a compound described herein (the“active ingredient”) into association with a carrier and/or one or moreother accessory ingredients, and then, if necessary and/or desirable,shaping and/or packaging the product into a desired single- ormulti-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient which would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition of the present disclosure will vary,depending upon the identity, size, and/or condition of the subjecttreated and further depending upon the route by which the composition isto be administered. By way of example, the composition may comprisebetween 0.1% and 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60),polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate(Span 40), sorbitan monostearate (Span 60], sorbitan tristearate (Span65), glyceryl monooleate, sorbitan monooleate (Span 80)),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor™) polyoxyethyleneethers, (e.g., polyoxyethylene lauryl ether (Brij 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimoniumbromide, cetylpyridinium chloride, benzalkonium chloride, docusatesodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol. Exemplary acidic preservatives include vitaminA, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid,dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

Other preservatives include tocopherol, tocopherol acetate, deteroximemesylate, cetrimide, butylated hydroxyanisol (BHA), butylatedhydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), sodium bisulfite, sodiummetabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus,Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, andEuxyl. In certain embodiments, the preservative is an anti-oxidant. Inother embodiments, the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, andwheat germ oils. Exemplary synthetic oils include, but are not limitedto, butyl stearate, caprylic triglyceride, capric triglyceride,cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate,mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixturesthereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the compoundsdescribed herein are mixed with solubilizing agents such as Cremophor™,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may comprise buffering agents.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type can be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active ingredient can be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets, and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a providedcompound may include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants and/or patches. Generally, the activeingredient is admixed under sterile conditions with a pharmaceuticallyacceptable carrier and/or any desired preservatives and/or buffers ascan be required. Additionally, the present disclosure encompasses theuse of transdermal patches, which often have the added advantage ofproviding controlled delivery of an active ingredient to the body. Suchdosage forms can be prepared, for example, by dissolving and/ordispensing the active ingredient in the proper medium. Alternatively oradditionally, the rate can be controlled by either providing a ratecontrolling membrane and/or by dispersing the active ingredient in apolymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices such as thosedescribed in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288;4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositionscan be administered by devices which limit the effective penetrationlength of a needle into the skin, such as those described in PCTpublication WO 99/34850 and functional equivalents thereof. Jetinjection devices which deliver liquid vaccines to the dermis via aliquid jet injector and/or via a needle which pierces the stratumcorneum and produces a jet which reaches the dermis are suitable. Jetinjection devices are described, for example, in U.S. Pat. Nos.5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189;5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335;5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880;4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballisticpowder/particle delivery devices which use compressed gas to acceleratevaccine in powder form through the outer layers of the skin to thedermis are suitable. Alternatively or additionally, conventionalsyringes can be used in the classical mantoux method of intradermaladministration.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi liquid preparations such as liniments,lotions, oil in water and/or water in oil emulsions such as creams,ointments and/or pastes, and/or solutions and/or suspensions.Topically-administrable formulations may, for example, comprise fromabout 1% to about 10% (w/w) active ingredient, although theconcentration of the active ingredient can be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition. Anotherformulation suitable for intranasal administration is a coarse powdercomprising the active ingredient and having an average particle fromabout 0.2 to 500 micrometers. Such a formulation is administered byrapid inhalation through the nasal passage from a container of thepowder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) and as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A provided pharmaceutical composition can be prepared,packaged, and/or sold in a formulation for buccal administration. Suchformulations may, for example, be in the form of tablets and/or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) active ingredient, the balance comprising an orallydissolvable and/or degradable composition and, optionally, one or moreof the additional ingredients described herein. Alternately,formulations for buccal administration may comprise a powder and/or anaerosolized and/or atomized solution and/or suspension comprising theactive ingredient. Such powdered, aerosolized, and/or aerosolizedformulations, when dispersed, may have an average particle and/ordroplet size in the range from about 0.1 to about 200 nanometers, andmay further comprise one or more of the additional ingredients describedherein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation for ophthalmic administration. Such formulationsmay, for example, be in the form of eye drops including, for example, a0.1/1.0% (w/w) solution and/or suspension of the active ingredient in anaqueous or oily liquid carrier. Such drops may further comprisebuffering agents, salts, and/or one or more other of the additionalingredients described herein. Other opthalmically-administrableformulations which are useful include those which comprise the activeingredient in microcrystalline form and/or in a liposomal preparation.Ear drops and/or eye drops are contemplated as being within the scope ofthis disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of provided compositionswill be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective dose level forany particular subject or organism will depend upon a variety of factorsincluding the disease, disorder, or condition being treated and theseverity of the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex and diet of the subject; the time of administration,route of administration, and rate of excretion of the specific activeingredient employed; the duration of the treatment; drugs used incombination or coincidental with the specific active ingredientemployed; and like factors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration).

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound(s), mode ofadministration, and the like. The desired dosage can be delivered threetimes a day, two times a day, once a day, every other day, every thirdday, every week, every two weeks, every three weeks, or every fourweeks. In certain embodiments, the desired dosage can be delivered usingmultiple administrations (e.g., two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, or moreadministrations).

In certain embodiments, an effective amount of a compound foradministration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound described herein may be administeredat dosage levels sufficient to deliver from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about mg/kg, from about 0.1 mg/kgto about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, orfrom about 1 mg/kg to about 25 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

In some embodiments, a compound described herein is administered one ormore times per day, for multiple days. In some embodiments, the dosingregimen is continued for days, weeks, months, or years.

It will be appreciated that dose ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

It will be also appreciated that a compound or composition, as describedherein, can be administered in combination with one or more additionaltherapeutically active agents. In certain embodiments, a compound orcomposition provided herein is administered in combination with one ormore additional therapeutically active agents that improve itsbioavailability, reduce and/or modify its metabolism, inhibit itsexcretion, and/or modify its distribution within the body. It will alsobe appreciated that the therapy employed may achieve a desired effectfor the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, priorto, or subsequent to, one or more additional therapeutically activeagents. In certain embodiments, the additional therapeutically activeagent is a compound of Formula (I). In certain embodiments, theadditional therapeutically active agent is not a compound of Formula(I). In general, each agent will be administered at a dose and/or on atime schedule determined for that agent. In will further be appreciatedthat the additional therapeutically active agent utilized in thiscombination can be administered together in a single composition oradministered separately in different compositions. The particularcombination to employ in a regimen will take into account compatibilityof a provided compound with the additional therapeutically active agentand/or the desired therapeutic effect to be achieved. In general, it isexpected that additional therapeutically active agents utilized incombination be utilized at levels that do not exceed the levels at whichthey are utilized individually. In some embodiments, the levels utilizedin combination will be lower than those utilized individually.

Exemplary additional therapeutically active agents include, but are notlimited to, small organic molecules such as drug compounds (e.g.,compounds approved by the U.S. Food and Drug Administration as providedin the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells.

Also encompassed by the present disclosure are kits (e.g.,pharmaceutical packs). The kits provided may comprise a providedpharmaceutical composition or compound and a container (e.g., a vial,ampule, bottle, syringe, and/or dispenser package, or other suitablecontainer). In some embodiments, provided kits may optionally furtherinclude a second container comprising a pharmaceutical excipient fordilution or suspension of a provided pharmaceutical composition orcompound. In some embodiments, a provided pharmaceutical composition orcompound provided in the container and the second container are combinedto form one unit dosage form. In some embodiments, a provided kitsfurther includes instructions for use.

Compounds and compositions described herein are generally useful for theinhibition of PRMT5. In some embodiments, methods of treatingPRMT5-mediated disorder in a subject are provided which compriseadministering an effective amount of a compound described herein (e.g.,a compound of Formula (I)), or a pharmaceutically acceptable saltthereof), to a subject in need of treatment. In certain embodiments, theeffective amount is a therapeutically effective amount. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the subject is suffering from aPRMT5-mediated disorder. In certain embodiments, the subject issusceptible to a PRMT5-mediated disorder.

As used herein, the term “PRMT5-mediated disorder” means any disease,disorder, or other pathological condition in which PRMT5 is known toplay a role. Accordingly, in some embodiments, the present disclosurerelates to treating or lessening the severity of one or more diseases inwhich PRMT5 is known to play a role.

In some embodiments, the present disclosure provides a method ofinhibiting PRMT5 comprising contacting PRMT5 with an effective amount ofa compound described herein (e.g., a compound of Formula (I)), or apharmaceutically acceptable salt thereof. The PRMT5 may be purified orcrude, and may be present in a cell, tissue, or subject. Thus, suchmethods encompass both inhibition of in vitro and in vivo PRMT5activity. In certain embodiments, the method is an in vitro method,e.g., such as an assay method. It will be understood by one of ordinaryskill in the art that inhibition of PRMT5 does not necessarily requirethat all of the PRMT5 be occupied by an inhibitor at once. Exemplarylevels of inhibition of PRMT5 include at least 10% inhibition, about 10%to about 25% inhibition, about 25% to about 50% inhibition, about 50% toabout 75% inhibition, at least 50% inhibition, at least 75% inhibition,about 80% inhibition, about 90% inhibition, and greater than 90%inhibition.

In some embodiments, provided is a method of inhibiting PRMT5 activityin a subject in need thereof comprising administering to the subject aneffective amount of a compound described herein (e.g., a compound ofFormula (I)), or a pharmaceutically acceptable salt thereof, or apharmaceutical composition thereof.

In certain embodiments, provided is a method of altering gene expressionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, provided is a method of altering transcriptionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, a method is provided of selecting a therapy fora subject having a disease associated with PRMT5-mediated disorder ormutation comprising the steps of determining the presence ofPRMT5-mediated disorder or gene mutation in the PRMT5 gene or andselecting, based on the presence of PRMT5-mediated disorder a genemutation in the PRMT5 gene a therapy that includes the administration ofa provided compound. In certain embodiments, the disease is cancer.

In certain embodiments, a method of treatment is provided for a subjectin need thereof comprising the steps of determining the presence ofPRMT5-mediated disorder or a gene mutation in the PRMT5 gene andtreating the subject in need thereof, based on the presence of aPRMT5-mediated disorder or gene mutation in the PRMT5 gene with atherapy that includes the administration of a provided compound. Incertain embodiments, the subject is a cancer patient.

In some embodiments, a provided compound is useful in treating aproliferative disorder, such as cancer, a benign neoplasm, an autoimmunedisease, or an inflammatory disease. For example, while not being boundto any particular mechanism, PRMT5 has been shown to be involved incyclin D1 dysregulated cancers. Increased PRMT5 activity mediates keyevents associated with cyclin D1-dependent neoplastic growth includingCUL4 repression, CDT1 overexpression, and DNA re-replication. Further,human cancers harboring mutations in Fbx4, the cyclin D1 E3 ligase,exhibit nuclear cyclin D1 accumulation and increased PRMT5 activity(Aggarwal et al., Cancer Cell. 2010 18(4):329-40). Additionally, PRMT5has also been implicated in accelerating cell cycle progression throughG1 phase and modulating regulators of G1; for example, PRMT5 mayupregulate cyclin-dependent kinase (CDK)₄, CDK6, and cyclins D1, D2 andE1. Moreover, PRMT5 may activate phosphoinositide 3-kinase (PI3K)/AKTsignaling (Wei et al., Cancer Sci. 2012 103(9):1640-50). Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating the following non-limiting list of cancers: breast cancer,esophageal cancer, bladder cancer, lung cancer, hematopoietic cancer,lymphoma, medulloblastoma, rectum adenocarcinoma, colon adenocarcinoma,gastric cancer, pancreatic cancer, liver cancer, adenoid cysticcarcinoma, lung adenocarcinoma, head and neck squamous cell carcinoma,brain tumors, hepatocellular carcinoma, renal cell carcinoma, melanoma,oligodendroglioma, ovarian clear cell carcinoma, and ovarian serouscystadenocarcinoma.

In some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating prostate cancer and lung cancer, in which PRMT5 hasbeen shown to play a role (Gu et al., PLoS One 2012; 7(8):e44033; Gu etal., Biochem. J. (2012) 446 (235-241)). In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of cancer. In some embodiments, a providedcompound is administered in combination with other compounds, drugs, ortherapeutics to treat cancer.

In some embodiments, compounds described herein are useful for treatinga cancer including, but not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma),familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g.,stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head andneck cancer (e.g., head and neck squamous cell carcinoma, oral cancer(e.g., oral squamous cell carcinoma (OSCC), throat cancer (e.g.,laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer,oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such asacute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acutemyelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma suchas Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkinlymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma(DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (i.e., “Waldenstrom's macroglobulinemia”), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma); a mixture of one or more leukemia/lymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease),hemangioblastoma, inflammatory myofibroblastic tumors, immunocyticamyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor,renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC),malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, smallcell lung cancer (SCLC), non-small cell lung cancer (NSCLC),adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g.,systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma,myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocyticleukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilicsyndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis(NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g.,gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoidtumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarianembryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma,pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductalpapillary mucinous neoplasm (IPMN), Islet cell tumors), penile cancer(e.g., Paget's disease of the penis and scrotum), pinealoma, primitiveneuroectodermal tumor (PNT), prostate cancer (e.g., prostateadenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer,skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA),melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g.,appendix cancer), soft tissue sarcoma (e.g., malignant fibroushistiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous glandcarcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g.,seminoma, testicular embryonal carcinoma), thyroid cancer (e.g.,papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC),medullary thyroid cancer), urethral cancer, vaginal cancer and vulvarcancer (e.g., Paget's disease of the vulva).

In some embodiments, a provided compound is useful in treating ametabolic disorder, such as diabetes or obesity. For example, while notbeing bound to any particular mechanism, a role for PRMT5 has beenrecognized in adipogenesis. Inhibition of PRMT5 expression in multiplecell culture models for adipogenesis prevented the activation ofadipogenic genes, while overexpression of PRMT5 enhanced adipogenic geneexpression and differentiation (LeBlanc et al., Mol. Endocrinol. 2012April; 26(4):583-97). Additionally, it has been shown that adipogenesisplays a pivotal role in the etiology and progression of diabetes andobesity (Camp et al., Trends Mol. Med. 2002 September; 8(9):442-7). Thusin some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating diabetes and/or obesity.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, diabetes. Insome embodiments, the diabetes is Type 1 diabetes. In some embodiments,the diabetes is Type 2 diabetes. In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of, obesity. In some embodiments, a providedcompound is useful to make a subject lose weight. In some embodiments, aprovided compound could be used in combination with other compounds,drugs, or therapeutics, such as metformin and insulin, to treat diabetesand/or obesity.

In some embodiments, a provided compound is useful in treating a blooddisorder, e.g., a hemoglobinopathy, such as sickle cell disease orβ-thalassemia. For example, while not being bound to any particularmechanism, PRMT5 is a known repressor of γ-globin gene expression, andincreased fetal γ-globin (HbF) levels in adulthood are associated withsymptomatic amelioration in sickle cell disease and β-thalassemia (Xu etal., Haematologica. 2012 November; 97(11):1632-40). Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating a blood disorder, such as a hemoglobinopathy such as sicklecell disease or β-thalassemia.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, sickle celldisease. In some embodiments, a provided compound is useful to delay theonset of, slow the progression of, or ameliorate the symptoms of,β-thalassemia. In some embodiments, a provided compound could be used incombination with other compounds, drugs, or therapeutics, to treat ahemoglobinopathy such as sickle cell disease or β-thalassemia.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 1. Compound B can be prepared via ringopening of a chiral or racemic epoxide group. This amino alcoholintermediate can be coupled to form an amide via normal amide couplingmethodology using a carboxylic acid A wherein Z is hydrogen or viaamination of an ester of intermediate A when Z is an optionallysubstituted aliphatic group.

For example, exemplary Schemes 2 and 3 show such couplings.

In some embodiments, an amide coupling step can be used to provide a keyintermediate for further synthesis, as shown, for example, in exemplaryScheme 4.

EXAMPLES

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Synthetic Methods Intermediate Synthesis2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1,2,3,4-tetrahydroisoquinoline (15 g, 0.11 mol) in MeCN(100 mL) was added K₂CO₃ (30.7 g, 0.23 mol) at 0° C.2-(bromomethyl)oxirane (17 g, 0.12 mol) was added to the reaction after1 h. The solution was stirred at 22° C. for 16 h at which time thesolids were filtered and washed with MeCN. The solution was concentratedand the residue was used in the next step without further purification(17 g, Yield 78%). LCMS (m/z): 190.1 (M+1).

1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline(17 g, 0.09 mol) in EtOH (300 mL) at −78° C. was slowly bubbled NH₃ (g).The reaction mixture was then sealed and heated at 80° C. for 3 h. Thereaction mixture was concentrated and the crude product was used in nextstep without further purification (18 g, Yield 96%). LCMS (m/z): 207.1(M+1).

(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1,2,3,4-tetrahydroisoquinoline (10 g, 0.15 mol) in THF(100 mL) at 0° C. was added KF (22 g, 0.3 mmol). After 1 h,(S)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (21.4 g, 0.17 mmol) wasadded and the resulting solution was stirred at 22° C. for 16 h. Thesolid was removed by filtration and washed with THF. The solution wasconcentrated and the crude compound was used for next step withoutfurther purification (15 g, Yield 53%). LCMS (m/z): 190.1 (M+1).

(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (15 g, 0.08mol) in EtOH (100 mL) at −78° C. was slowly bubbled NH₃ (g). Thereaction mixture was then sealed and heated at 80° C. for 3 h. Thereaction mixture was concentrated and the crude product was used in nextstep without further purification (15 g, Yield 92%). LCMS (m/z): 207.1(M+1).

Alternative Synthesis of(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1,2,3,4-tetrahydroisoquinoline (1 g, 7.52 mmol) in MeOH(40 mL) was added K₂CO₃ (5.19 g, 37.6 mmol) under 0° C. After stirringfor 30 minutes, (R)-2-(chloromethyl) oxirane (0.692 g, 7.52 mmol) wasadded the reaction. The mixture was then stirred at 0° C. overnightbefore filtration and washing of the solid by with MeOH. The solutionwas concentrated and the residue purified by column separation to givethe title compound as a colorless oil (70% purity). This crude was useddirectly in the next step. LCMS (m/z): 190.1 (M+1).

Alternative Synthesis of(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of(R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (200 mg, 5.2mmol) in EtOH (20 mL) was added NH₄OH (600 mg, 35.2 mmol) at −78° C. Thereaction mixture was then warmed and heated at 100° C. for 3 h in a sealtube. The reaction mixture was concentrated and the crude product wasused in next step without further purification. LCMS (m/z): 207.1 (M+1).

(S)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1,2,3,4-tetrahydroisoquinoline (5 g, 7.52 mmol) in THF(100 mL) was added KF (8.57 g, 150.4 mmol) at 0° C.(R)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (10.7 g, 41.4 mmol) wasadded to the reaction in 1 h. The solution was stirred at roomtemperature overnight. The solid was removed by filtration and washedwith THF. The solution was then concentrated and the residue used fornext step without further purification (11.3 g Yield 80%). LCMS (m/z):190.1 (M+1).

(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol

To a solution of(S)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (2.2 g, 0.012mol) in EtOH (30 mL), NH₃ was bubbled to the solution under −78° C. Thereaction mixture was then sealed and heated at 80° C. for 3 h. AfterLCMS indicated completion of the reaction, the mixture was concentratedand the crude product was used in next step without further purification(2.2 g, Yield 90%). LCMS (m/z): 207.1 (M+1).

Compound 1N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxyacetamide

To a stirred mixture of 2-phenoxyacetic acid (100 mg, 0.658 mmol) in DCM(10 mL) was added TEA (200 mg, 1.98 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (135 mg, 0.658mmol) and HATU (250 mg, 0.658 mmol). The mixture was stirred at 25° C.for 16 hours then quenched with water (20 mL) and extracted with DCM(3×20 mL). The combined extracts were washed with brine (20 mL), driedover anhydrous Na₂SO₄ and concentrated. The residue was then purified byprep-HPLC to afford the title compound (76 mg, 34% yield). ¹H NMR (400MHz, CDCl₃): δ 7.31-7.27 (m, 2H), 7.14-7.08 (m, 3H), 7.05-6.98 (m, 2H),6.93 (d, J=8.0 Hz, 2H), 4.54 (s, 2H), 4.55-4.52 (m, 1H), 4.42 (s, 2H),4.06-4.00 (m, 1H), 3.71 (s, 2H), 3.48-3.38 (m, 2H), 2.91 (d, J=5.6 Hz,2H), 2.84 (d, J=5.6 Hz, 2H), 2.60 (d, J=6.8 Hz, 2H) ppm. LCMS (m/z):341.2 [M+H]⁺.

Compound 6N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)acetamide

Step 1: 2-(quinolin-8-yloxy)acetic acid

To a stirred mixture of quinolin-8-ol (500 mg, 3.45 mmol) in MeCN (5 mL)was added ethyl bromoacetate (687 mg, 4.14 mmol) and K₂CO₃ (952 mg, 6.90mmol). The mixture was stirred at 80° C. for 4 hours until TLC analysisshowed completion of the reaction. The mixture was filtered and thefiltrate concentrated. NaOH (276 mg, 6.90 mmol) and water:EtOH (1:1, 10mL) was then added to the residue and the resulting mixture stirred at50° C. for 4 hours. After cooling, the mixture was acidified by additionof 1M HCl to pH 3 and then extracted with ethyl acetate (2×30 mL). Thecombined organic layers were washed with brine (30 mL), dried overanhydrous Na₂SO₄ and concentrated to yield the crude target productwhich was used directly for the next step.

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)acetamide

To a stirred mixture of 2-(quinolin-8-yloxy)acetic acid (100 mg, 0.492mmol) in DMF (5 mL) was added DIEA (95 mg, 0.738 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (100 mg, 0.492mmol) and BOP-Cl (151 mg, 0.591 mmol). The mixture was stirred at 25° C.for 48 hours then the reaction mixture was quenched by addition of water(20 mL) and extracted with DCM (3×20 mL). The combined organic extractswere washed with brine (20 mL), dried over anhydrous Na₂SO₄ andconcentrated. The residue was then purified by prep-HPLC to afford thedesired product (8 mg, Yield: 4%). ¹H NMR (400 MHz, MeOD) δ=8.91 (d,J=4.3 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 7.69-7.54 (m, 3H), 7.30 (d, J=7.3Hz, 1H), 7.12-6.96 (m, 4H), 4.78 (s, 2H), 4.18-4.07 (m, 1H), 3.71 (s,2H), 3.60-3.49 (m, 1H), 3.48-3.40 (m, 1H), 2.89 (d, J=5.8 Hz, 2H), 2.84(d, J=4.8 Hz, 2H), 2.65-2.61 (m, 2H). LCMS (m/z): 392.2 (M+1).

Compound 7N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-fluorophenoxy)acetamide

Step 1: 2-(3-fluorophenoxy)acetic acid

To a stirred mixture of 3-fluorophenol (100 mg, 0.893 mmol) in MeCN (5mL) was added ethyl bromoacetate (222 mg, 1.34 mmol) and K₂CO₃ (369 mg,2.68 mmol). The mixture was stirred at 80° C. for 4 hours. The mixturewas then filtered and the filtrate concentrated. NaOH (71 mg, 1.79 mmol)and water:EtOH (1:1, 10 mL) was added to the residue and the mixturestirred at 50° C. for 4 hours. The mixture was acidified by adding 1MHCl, and then extracted with ethyl acetate (2×30 mL). The combinedextracts were washed with brine (30 mL), dried over anhydrous Na₂SO₄ andconcentrated. The residue was directly for the next step. LCMS (m/z):171.0 (M+1).

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-fluorophenoxy)acetamide

To a stirred mixture of 2-(3-fluorophenoxy)acetic acid (252 mg, 0.893mmol) in DCM (5 mL) was added DIEA (173 mg, 1.34 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (183 mg, 0.893mmol), and Bop-Cl (273 mg, 1.07 mmol). The mixture was stirred at 25° C.for 16 hours then quenched with water (20 mL), extracted with DCM (3×20mL). The combined extracts were washed with brine (20 mL), dried overanhydrous Na₂SO₄ and concentrated. The residue was then purified byprep-HPLC to afford the product (18 mg, Yield 5.6%). ¹H NMR (400 MHz,METHANOL-d₄) δ=7.26-7.29 (m, 1H), 7.13-7.02 (m, 4H), 6.76-6.69 (m, 3H),4.55 (s, 2H), 4.06-4.00 (m, 1H), 3.71 (s, 1H), 3.47-3.31 (m, 2H),2.91-2.90 (m, 2H), 2.86-2.77 (m, 2H), 2.61 (d, J=6.0 Hz, 2H). LCMS(m/z): 359.1 (M+1).

Compound 82-(3-cyanophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

Step 1: 2-(3-cyanophenoxy)acetic acid

To a stirred mixture of 3-hydroxybenzonitrile (100 mg, 0.840 mmol) inMeCN (5 mL) was added ethyl bromoacetate (209 mg, 1.26 mmol) and K₂CO₃(350 mg, 2.52 mmol). The mixture was stirred at 80° C. for 4 hours untilTLC showed completion of the reaction. The mixture was filtered and thefiltrate concentrated. NaOH (67 mg, 1.68 mmol) and water:EtOH (1:1, 10mL) was added to the residue and the mixture stirred at 50° C. for 4hours. After cooling, the mixture was acidified by 1M HCl, extractedwith ethyl acetate (2×30 mL). The combined extracts were washed withbrine (30 mL), dried over anhydrous Na₂SO₄ and concentrated. The residuewas used directly in the next step. LCMS (m/z): 178.0 (M+1).

Step 2:2-(3-cyanophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

To a stirred mixture of 2-(3-cyanophenoxy)acetic acid (100 mg, 0.565mmol) in DCM (5 mL) was added DIEA (109 mg, 0.85 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (116 mg, 0.565mmol) and BopCl (173 mg, 0.678 mmol). The resulting mixture was stirredat 25° C. for 16 hours until LCMS showed the completion of the reaction.The reaction mixture was quenched by addition of water (20 mL) thenextracted with DCM (3×20 mL). The combined organic extracts were washedwith brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated. Theresidue was purified by prep-HPLC to afford the desired final product(24 mg, Yield 12%). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.48 (dd, J₁=J₂=7.6Hz, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.25-7.21 (m, 2H), 7.13-7.03 (m, 4H),4.60 (s, 2H), 4.04-4.00 (m, 1H), 3.71 (s, 2H), 3.44 (d, J=6.0 Hz, 2H),2.93-2.80 (m, 4H), 2.62 (d, J=5.6 Hz, 2H). LCMS (m/z): 366.1 (M+1).

Compound 9N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(p-tolyloxy)acetamide

Step 1: ethyl 2-(p-tolyloxy)acetate

To a mixture of p-cresol (500 mg, 4.63 mmol) and ethyl 2-bromoacetate(928 mg, 5.56 mmol) in CH₃CN (10 mL) was added K₂CO₃ (3 g, 21.7 mmol).The reaction mixture was stirred at 80° C. for 4 h. The solid wasremoved by filtration and the filtrate was concentrated to give thetitle compound which was used in the next step without furtherpurification.

Step 2: 2-(p-tolyloxy)acetic acid

To a solution of ethyl 2-(p-tolyloxy)acetate (200 mg, 1 mmol) in EtOH(10 ml) was added 10% NaOH solution (10 ml) at 26° C. The mixture wasstirred for 30 min, concentrated then water (20 mL) added to it beforewashing with ethyl acetate (2×20 mL). The aqueous layer was acidifiedwith 2N HCL until pH 3 and extracted with EA (2×20 ml). The organiclayer was washed with brine (30 mL), dried over Na₂SO₄ and concentratedto give the title compound which was used in next step without furtherpurification.

Step3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(p-tolyloxy)acetamide

A mixture of compound 2-(p-tolyloxy)acetic acid (100 mg, 0.60 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (124 mg, 0.60mmol), BOP-Cl (183 mg, 0.72 mmol) and DIPEA (1 mL) in DCM (10 mL) wasstirred at room temperature for 4 h. The solvent was removed byconcentration and the crude product was purified by pre-HPLC to give thetitle compound (27.8 mg, yield 13.1%). ¹H NMR (500 MHz, MeOD): δ7.32-7.25 (m, 3H), 7.20 (d, J=7.2 Hz, 1H), 7.12 (d, J=8.4 Hz, 2H), 6.90(d, J=8.8 Hz, 2H), 4.65-4.52 (br.s, 1H), 4.52 (s, 2H), 4.46-4.30 (br.s,1H), 4.30-4.24 (m, 1H), 3.85-3.70 (br.s, 1H), 3.43 (d, J=5.6 Hz, 1H),3.26-3.17 (m, 4H), 2.26 (s, 3H) ppm; ESI-MS (m/z): 355.2 [M+1]⁺.

Compound 12N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methoxyphenoxy)acetamide

Step 1: ethyl 2-(4-methoxyphenoxy)acetate

To a solution of 4-methoxyphenol (500 mg, 4.03 mmol) in CH₃CN (10 mL)was added ethyl 2-bromo-2-methylpropanoate (807 mg, 4.84 mmol) and K₂CO₃(3 g, 21.7 mmol) at 25° C. The mixture was refluxed for 16 h. Themixture was then diluted with water (100 mL), extracted with ethylacetate (2×50 mL) and the combined organic layers washed with brine (30mL), dried over Na₂SO₄ and concentrated to give the title compound whichwas used in next step without further purification.

Step 2: 2-(4-methoxyphenoxy)acetic acid

To a solution of ethyl 2-methyl-2-phenoxypropanoate (210 mg, 1 mmol) inEtOH (10 ml) was added 10% NaOH aqueous solution (10 mL) at 26° C. Themixture was stirred for 30 min and then concentrated before the additionof water (20 mL) and washing with ethyl acetate (2×20 mL). The aqueouslayer was acidified with 2N HCL until pH 3 and extracted with ethylacetate (2×20 ml). The organic layer was washed with brine (30 mL),dried over Na₂SO₄ and concentrated to give the title compound which wasused in next step without further purification.

Step 3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methoxyphenoxy)acetamide

A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100 mg, 0.55mmol), 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (113.7 mg,0.55 mmol), BOP-Cl (177.2 mg, 0.696 mmol) and DIPEA (1 mL) in DCM (10mL) was stirred at room temperature for 4 h. The solvent was removed byconcentration and the crude product was purified by pre-HPLC to give thetitle compound (13.3 mg, yield 6.5%). ¹H NMR (500 MHz, MeOD): δ7.11-7.06 (m, 3H), 7.02-7.00 (m, 1H), 6.90-6.60 (m, 4H), 4.45 (s, 2H),4.01-3.98 (m, 1H), 3.73 (s, 3H), 3.66 (s, 1H), 3.41-3.39 (m, 2H),2.90-2.87 (m, 2H), 2.81-2.76 (m, 2H), 2.55 (d, J=6.0 Hz, 2H) ppm; ESI-MS(m/z): 371.2 [M+1]⁺.

Compound 15N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(o-tolyloxy)acetamide

Step 1: 2-(o-tolyloxy)acetic acid

To a stirred solution of o-cresol (200 mg, 1.85 mmol) in MeCN (5 mL) wasadded ethyl bromoacetate (461 mg, 2.78 mmol) and K₂CO₃ (766 mg, 5.55mmol). The mixture was stirred at 80° C. for 4 hours. The mixture wasfiltered and the filtrate concentrated. NaOH (150 mg, 3.70 mmol) andH₂O/EtOH (1:1, 10 mL) was then added to the mixture and the mixturestirred at 50° C. for 4 hours. After cooling, the mixture was acidifiedby adding 1M HCl then extracted with ethyl acetate (2×30 mL). Thecombined organic extracts were washed with brine (30 mL), dried overanhydrous Na₂SO₄ and concentrated. The residue was used directly in thenext step without further purification.

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(o-tolyloxy)acetamide

To a stirred mixture of 2-(o-tolyloxy)acetic acid (100 mg, 0.60 mmol) inDCM (5 mL) was added DIEA (116 mg, 0.90 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (124 mg, 0.60mmol) and Bop-Cl (183 mg, 0.72 mmol). The mixture was stirred at 25° C.for 16 hours then quenched by addition of water (20 mL). The resultingmixture was extracted with DCM (3×20 mL). The combined extracts werewashed with brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated.The residue was purified by prep-HPLC to afford the product (79 mg,Yield 37%). ¹H NMR (400 MHz, MeOD) δ 7.14-7.01 (m, 6H), 6.95-6.82 (m,2H), 4.55 (s, 2H), 4.04-4.01 (m, 1H), 3.72-3.63 (m, 2H), 3.55-3.46 (m,1H), 3.42-3.34 (m, 1H), 2.93-2.84 (m, 2H), 2.83-2.74 (m, 2H), 2.57 (d,J=6.3 Hz, 2H), 2.27 (s, 3H). LCMS (m/z): 355.1 (M+1).

Compound 19N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxypropanamide

Step 1: ethyl 2-phenoxypropanoate

To a solution of NaH (765 mg, 31.89 mmol) in DMF (10 mL) was addedphenol (1 g, 10.63 mmol) at 25° C. The mixture was heated at refluxtemperature for 15 min, and then ethyl 2-bromopropanoate (2.3 g, 12.75mmol) was added. The resulting mixture was stirred at 25° C. for another16 h before quenching with water (50 mL). The mixture was extracted withethyll acetate (3×20 mL). The combined organic layers were washed withbrine (30 mL), dried over Na₂SO₄ and concentrated to give the titlecompound (1.8 g, 85.7%) as colorless oil which was used in next stepwithout further purification.

Step 2: 2-phenoxypropanoic acid

To a solution of ethyl 2-phenoxypropanoate (1.8 g, 0.9 mmol) in EtOH (16ml) was added a solution of NaOH (0.44 g, 1.1 mmol) in H₂O (4 ml) at 25°C. The mixture was stirred for 30 min before being concentrated. Theresidue had water (20 mL) added and washed with ethyl acetate (2×20 mL).The aqueous layer was acidified with 2N HCL until pH 3 and extractedwith ethyl acetate (2×20 mL). The combined organic layers were washedwith brine (30 mL), dried over Na₂SO₄ and concentrated to give the titlecompound (1.1 g, 73.3%) as a white solid which was used in next stepwithout further purification.

Step 3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-phenoxypropanamide

To a solution of 2-phenoxypropanoic acid (200 mg, 1.2 mmol) in DMF (4ml) was added TEA 364 mg, 3.6 mmol), HOBt (243 mg, 1.8 mmol), EDCI (346mg, 1.8 mmol) and 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol(297 mg, 1.44 mmol) at 24° C. The reaction mixture was stirred for 16 huntil TLC showed completion of the reaction. After evaporation of thesolvent, the residue was purified by prep-HPLC separation to give thetitle compound as the formate salt (34 mg, 8%). ¹H NMR (500 MHz, MeOD):δ 8.40 (s, 1H), 7.34-7.25 (m, 5H), 7.17 (d, J=7.2 Hz, 1H), 7.00-6.96 (m,3H), 4.78-4.76 (m, 1H), 4.26-4.11 (m, 3H), 3.43-3.33 (m, 4H), 3.14-3.12(m, 2H), 3.08-3.02 (m, 1H), 2.95-2.90 (m, 1H), 1.57 (d, J=6.4 Hz, 3H)ppm; ESI-MS (m/z): 354.2 [M+1]⁺.

Compound 20N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-methoxyphenoxy)acetamide

Step 1: 2-(3-methoxyphenoxy)acetic acid

To a stirred mixture of 3-methoxyphenol (200 mg, 1.61 mmol) in MeCN (5mL) was added ethyl bromoacetate (402 mg, 2.42 mmol) and K₂CO₃ (672 mg,4.83 mmol). The mixture was stirred at 80° C. for 4 hours, filtered andthe filtrate concentrated. NaOH (129 mg, 3.22 mmol) and H2O/EtOH (1:1,10 mL) was added to the mixture. The reaction mixture was stirred at 50°C. for 4 hours then acidified by 1M HCl and then extracted with ethylacetate (2×30 mL). The combined organic extracts were washed with brine(30 mL), dried over anhydrous Na₂SO₄ and concentrated with the residueused directly for the next step. LCMS (m/z): 183.0 (M+1).

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-methoxyphenoxy)acetamide

To a stirred mixture of 2-(3-methoxyphenoxy)acetic acid (100 mg, 0.549mmol) in DCM (5 mL) was added DIEA (106 mg, 0.824 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (113 mg, 0.549mmol) and Bop-Cl (168 mg, 0.66 mmol). The mixture was stirred at 25° C.for 16 hours then the reaction mixture was quenched with water (20 mL),extracted with DCM (3×20 mL). The combined extracts were washed withbrine (20 mL), dried over anhydrous Na₂SO₄ and concentrated. The residuewas purified by prep-HPLC to afford the desired product (61 mg, Yield32%). ¹H NMR (400 MHz, MeOD) δ 7.21 (dd, J₁=J₂=8.0 Hz, 1H), 7.17-7.03(m, 4H), 6.59-6.51 (m, 3H), 4.52 (s, 2H), 4.05-4.01 (m, 1H), 3.77 (s,3H), 3.73 (d, J=2.8 Hz, 2H), 3.47-3.37 (m, 2H), 2.92 (d, J=5.2 Hz, 2H),2.87 (d, J=5.2 Hz, 2H), 2.61 (d, J=6.0 Hz, 2H). LCMS (m/z): 371.1 (M+1).

Compound 212-(4-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

Step 1: ethyl 2-(4-acetamidophenoxy)acetate

To a solution of N-(4-hydroxyphenyl)acetamide (500 mg, 3.31 mmol) inCH₃CN (10 mL) was added ethyl 2-bromo-2-methylpropanoate (663 mg, 3.97mmol) and K₂CO₃ (3 g, 21.7 mmol) at 25° C. The mixture was heated atreflux for 16 h. The mixture had water (100 mL) added and extracted withethyl acetate (2×50 mL). The combined organic layers were washed withbrine (30 mL), dried over Na₂SO₄ and concentrated to give the titlecompound which was used in next step without further purification.

Step 2: 2-(4-acetamidophenoxy)acetic acid

To a solution of ethyl 2-(4-acetamidophenoxy)acetate (237 mg, 1 mmol) inEtOH (10 ml) was added 10% NaOH aqueous solution (10 mL) at 26° C. Themixture was stirred for 30 min and then concentrated. The residue wasdiluted with water (20 mL) and washed ethyl acetate (2×20 mL). Theaqueous layer was then acidified with 2N HCL until pH 3 and extractedwith ethyl acetate (2×20 ml). The combined organic layers were washedwith brine (30 mL), dried over Na₂SO₄ and concentrated to give the titlecompound which was used in next step without further purification.

Step 3:2-(4-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100 mg, 0.51mmol), 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg,0.51 mmol), BOP-Cl (156 mg, 0.61 mmol) and DIPEA (1 mL) in DCM (10 mL)was stirred at room temperature for 4 h. The solvent was removed byconcentration and the crude product was purified by pre-HPLC to give thetitle compound (7.8 mg, yield: 3.8%). ¹H NMR (500 MHz, MeOD): δ 7.45 (d,d, J=8.8 Hz, 2H), 7.11-7.05 (m, 3H), 7.02-7.00 (m, 1H), 6.88 (d, J=8.8Hz, 2H), 4.50 (s, 2H), 4.01-3.98 (m, 1H), 3.66 (d, J=3.2 Hz, 2H), 3.41(dd, J=0.8, 6.0 Hz, 2H), 2.89-2.87 (m, 2H), 2.80-2.76 (m, 2H), 2.55 (d,J=6.0 Hz, 2H), 2.08 (s, 3H) ppm; ESI-MS (m/z): 398.2 [M+1]⁺.

Compound 23N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-indazol-6-yl)oxy)acetamide

Step 1: 1-methyl-1H-indazol-6-amine

To a solution of 1-methyl-6-nitro-1H-indazole (1.2 g, 0.7 mmol) in EtOH(50 ml) and THF (15 ml) was added PtO₂ (125 mg) at 26° C. The mixturewas stirred for 1.5 h at 26° C. under a H₂ atmosphere at 30 Psi. Oncethe reaction was complete by TLC analysis, the mixture was filtered andthe filtrate concentrated to give the target crude product as a whitesolid which was used in the next step without further purification (1.0g, Yield 90%). LCMS (m/z): 148.1 (M+1).

Step 2: 1-methyl-1H-indazol-6-ol

To a solution of 1-methyl-1H-indazol-6-amine (300 mg, 2.04 mmol) inH₂O/H₂SO₄=1:1 (5 ml) was added NaNO₂ (141 mg, 2.04 mmol) at 0° C. Themixture was then stirred for 2 h at 25° C. before being added to water(0.5 ml) and stirred for a further 2 h at 120° C. Once the reaction wascomplete by TLC, the mixture was treated with NaHCO₃ until pH=7. Themixture was then extracted with ethyl acetate (2×10 ml) and the organiclayer washed with brine (20 ml), dried over Na₂SO₄ and concentrated togive 1-methyl-1H-indazol-6-ol as a red solid which was used in the nextstep without further purification (300 mg, 99.0%). LCMS (m/z):149.1(M+1).

Step 3: 2-((1-methyl-1H-indazol-6-yl)oxy)acetic acid

To a solution of NaH (146 mg, 6.06 mmol) in DMF (3 mL) was added1-methyl-1H-indazol-6-ol (300 mg, 2.02 mmol) at 25° C. After stirringfor 5 minutes, ethyl 2-bromoacetate (406 mg, 2.43 mmol) was added andstirred for 16 h at 25° C. The mixture was then diluted with water (50mL) and washed with ethyl acetate (2×20 mL). The water layer was thenacidified by adding with 2N HCL until pH3 and then extracted withethylacetate (2×20 ml). The combined organic layers were washed withbrine (30 mL), dried over Na₂SO₄ and concentrated to give the crudeproduct (300 mg, 63.4%) as colorless oil. It was used in next stepwithout further purification. LCMS (m/z): 207.1 (M+1).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-indazol-6-yl)oxy)acetamide

To a solution of 2-((1-methyl-1H-indazol-6-yl)oxy)acetic acid (200 mg,0.97 mmol) in DMF (4 ml) was added TEA (294 mg, 2.91 mmol), HOBt (196mg, 1.45 mmol), EDCI (278 mg, 1.45 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (240 mg, 1.2mmol) at 27° C. The reaction mixture was stirred for 16 h at 27° C. Oncethe reaction was complete and evaporation of the solvent, the mixturewas purified by preparative HPLC to give the formate salt of the titlecompound (26 mg, 11.9%) as a white solid. ¹H NMR (400 MHz, MeOD) δ 7.91(s, 1H), 7.67 (d, J=8.8 Hz, 1H), 7.27-7.13 (m, 4H), 7.02 (s, 1H), 6.95(dd, J₁=8.8 Hz, J₂=2.0 Hz, 1H), 4.68 (s, 2H), 4.26-4.25 (m, 3H), 4.02(s, 3H), 3.47-3.33 (m, 4H), 3.14-3.06 (m, 4H). LCMS (m/z): 395.2 (M+1).

Compound 242-(cyclohexyloxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

Step 1: 2-(cyclohexyloxy)acetic acid

To a solution of compound NaH (719 mg, 29.94 mmol) in DMF (10 mL) wasadded cyclohexanol (1 g, 9.98 mmol) at 0° C. After stirring for 5minutes, ethyl 2-bromoacetate (2 g, 11.98 mmol) was added and themixture stirred for another 16 h. Once complete, the mixture was treatedwith water (50 mL) and washed with ethyl acetate (2×20 mL). The waterlayer was treat with 2N HCL until pH 3. The water layer was extractedwith ethyl acetate (2×20 ml) and the combined organic layers washed withbrine (30 mL), dried over Na₂SO₄ and concentrated to give the desiredproduct (500 mg, 27.8%) as colorless oil which was used in next stepwithout further purification.

Step 2:2-(cyclohexyloxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

To a solution of 2-(cyclohexyloxy)acetic acid (100 mg, 0.632 mmol) inDMF (3 ml) was added TEA (191 mg, 1.896 mmol), HOBT (128 mg, 0.948mmol), EDCI (182 mg, 0.948 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (156 mg, 0.758mmol) at 27° C. The mixture was stirred for 16 h until the reaction wascomplete. After evaporation of the solvent, the residue was purified byprep-HPLC to afford the title compound as the formate salt (26 mg, Yield11.9%). ¹H NMR (400 MHz, MeOD): δ 8.48 (s, 1H), 7.27-7.22 (m, 3H),7.16-7.15 (m, 1H), 4.23-4.18 (m, 3H), 4.00 (s, 2H), 3.44-3.33 (m, 3H),3.13-3.02 (m, 4H), 1.95-1.13 (m, 2H), 1.78-1.76 (m, 2H), 1.63-1.61 (m,1H), 1.37-1.27 (m, 5H) ppm; ESI-MS (m/z): 469.3 [M+1]⁺.

Compound 25N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-phenoxypropanamide

Step 1: ethyl 2-methyl-2-phenoxypropanoate

To a solution of phenol (2 g, 21.25 mmol) in CH₃CN (50 mL) was addedethyl 2-bromo-2-methylpropanoate (5 g, 25.5 mmol) and Cs₂CO₃ (20 g,63.75 mmol) at 25° C. The mixture was heated at reflux for 16 h thenwater (100 mL) added and extracted with ethyl acetate (2×50 mL). Theorganic layer was washed with brine (30 mL), dried over Na₂SO₄ andconcentrated to give the title compound (2.1 g, 47.7%) as colorless oilwhich was used in next step without further purification.

Step 2: 2-methyl-2-phenoxypropanoic acid

To a solution of ethyl 2-methyl-2-phenoxypropanoate (2.0 g, 9.6 mmol) inEtOH (16 ml) was added a solution of NaOH (0.46 g, 11.5 mmol) in H₂O (4ml) at 26° C. The mixture was stirred for 30 min then concentrated.Water was added (20 mL) and washed with ethyl acetate (2×20 mL). Theaqueous layer was acidified with 2N HCL until pH 3 and extracted withethyl acetate (2×20 ml). The combined organic layers were washed withbrine (30 mL), dried over Na₂SO₄ and concentrated to give the titlecompound (1.6 g, 94.1%) as a white solid which was used in next stepwithout further purification.

Step 3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-phenoxypropanamide

To a solution of compound 2-methyl-2-phenoxypropanoic acid (200 mg, 1.11mmol) in DMF (4 ml) was added TEA (336 mg, 3.33 mmol), HOBt (225 mg,1.66 mmol), EDCI (320 mg, 1.66 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (320 mg, 1.33mmol) at 24° C. The reaction mixture was stirred for 16 h at 24° C.After evaporation of the solvent, the residue was purified by prep-HPLCseparation to give the title compound as the formate salt (33 mg, 8%).¹H NMR (500 MHz, MeOD): δ 8.40 (s, 1H), 7.32-7.25 (m, 5H), 7.19 (d,J=6.8 Hz, 1H), 7.09 (t, J=7.6 Hz, 1H), 6.98-6.96 (m, 2H), 4.33 (s, 2H),4.29-4.22 (m, 1H), 3.49 (t, J=6.4 Hz, 2H), 3.42 (d, J=5.6 Hz, 2H),3.16-3.07 (m, 4H), 1.51 (s, 6H) ppm; ESI-MS (m/z): 369.5 [M+1]⁺.

Compound 28N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(methylsulfonamido)phenoxy)acetamide

MsCl (23 mg, 0.2 mmol) was added to a cooled 0° C. stirred solution of2-(3-aminophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide(71 mg, 0.2 mmol) in Et₃N (0.1 mL) and DCM (10 mL). After stirred for 2h, the solvent was removed by concentration. The residue was purified byprep-HPLC to afford the title compound. ¹H NMR (400 MHz, MeOD): δ7.26-7.22 (m, 1H), 7.11-7.01 (m, 4H), 6.91-6.85 (m, 2H), 6.71-6.68 (m,1H), 4.53 (s, 2H), 4.03-4.00 (m, 1H), 3.69 (s, 2H), 3.43-3.88 (m, 1H),3.13-3.12 (m, 1H), 2.96 (s, 3H), 2.89 (d, J=6 Hz, 1H), 2.82 (d, J=5.6Hz, 1H), 2.57 (t, J=6 Hz, 1H) ppm; ESI-MS (m/z): 434.1 [M+1]⁺.

Compound 30N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetamide

Step 1: 3-(methylamino)-4-nitrophenol

The solution of 3-fluoro-4-nitrophenol (1 g, 6.37 mmol) in aqueous MeNH₂solution (5 mL) was stirred at 85° C. for 5 h. After cooling to roomtemperature, the solution was diluted with water (30 mL) andconcentrated HCl added to adjust to pH 1. The resulting precipitate wascollected by filtration and the solid dried under vacuum to give thecrude product which was used without further purification (1.1 g, 95%yield). LCMS (m/z): 169.1 (M+1).

Step 2: 1-methyl-1H-benzo[d]imidazol-6-ol

Fe Powder (4.33 g, 77.4 mmol) was added to a solution of3-(methylamino)-4-nitrophenol (1.3 g, 7.74 mmol) in HCOOH (30 mL) andthe mixture heated to 100° C. for 16 h. After cooling to roomtemperature, MeOH (250 mL) was added to mixture and filtered over a padof Celite. The filtrate was collected, concentrated and the residuepurified by column chromatography to give the crude desired product (1.2g) and was used directly in the next step. LCMS (m/z): 149.06 (M+1).

Step 3: ethyl 2-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetate

The mixture of 1-methyl-1H-benzo[d]imidazol-6-ol (600 mg, 4.03 mmol),BrCH₂COOEt (372 mg, 4.03 mmol) and K₂CO₃ (1.1 g, 8.06 mmol) in DMF (8mL) was stirred at room temperature for 16 h. DCM (100 mL) and water(100 mL) was then added to the reaction and the organic layer washedwith water (50 mL), brine (50 mL) and dried over Na₂SO₄ before filteringand concentration to give the crude desired product (560 mg, Yield 60%).LCMS (m/z): 235.1 (M+1).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetamide

A neat solution of ethyl2-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)acetate (100 mg, 0.427 mmol)and 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (88 mg, 0.427mmol) was stirred at room temperature for 2 days until TLC showed thecompletion of the reaction. The solution concentrated and the residuepurified by prep-HPLC to give desired product as the TFA salt (19.1 mg,Yield 11.3%). ¹H NMR (400 MHz, MeOD). δ 9.21 (s, 1H), 7.80 (d, J=8.8 Hz,1H), 7.49 (s, 1H), 7.40-7.20 (m, 5H), 4.74 (s, 2H), 4.53 (br.s, 2H),4.37-4.32 (m, 1H), 4.11 (s, 3H), 3.68 (br.s, 2H), 3.53-3.12 (m, 6H).LCMS (m/z): 395.1 (M+1).

Compound 312-(3-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

Step 1: 2-(3-acetamidophenoxy)acetic acid

To a stirred mixture of N-(3-hydroxyphenyl)acetamide (300 mg, 2.0 mmol)in MeCN (5 mL) was added ethyl bromoacetate (500 mg, 3 mmol) and K₂CO₃(828 mg, 6 mmol). The mixture was stirred at 80° C. for 4 hours,filtered and the filtrate was concentrated. NaOH (80 mg, 2 mmol) andwater:EtOH (1:1, 10 mL) was added to the mixture. This mixture was thenstirred at 50° C. for 4 hours before being acidified by 1M HCl,extracted with ethyl acetate (2×30 mL). The combined extracts werewashed with brine (30 mL), dried over anhydrous Na₂SO₄ and concentratedwith the residue used directly for the next step. LCMS (m/z): 183.0(M+1).

Step 2:2-(3-acetamidophenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

To a stirred mixture of 2-(3-acetamidophenoxy)acetic acid (85 mg, 0.41mmol) in DCM (5 mL) was added TEA (0.5 mL),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (83.8 mg, 0.41mmol) and HATU (171 mg, 0.451 mmol). The mixture was stirred at 25° C.for 3 hours. The reaction mixture was quenched by addition of water (20mL) and extracted with DCM (3×20 mL) and the combined extracts washedwith brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated. Theresidue was purified by prep-HPLC to afford the title product (55 mg,Yield 35%). ¹H NMR (400 MHz, MeOD) δ=7.36 (s, 1H), 7.27-7.17 (m, 1H),7.19-7.01 (m, 6H), 6.68-6.66 (m, 1H), 4.53 (s, 2H), 4.03-4.00 (m, 1H),3.71-3.62 (m, 2H), 3.50-3.35 (m, 2H), 2.95-2.85 (m, 2H), 2.83-2.74 (m,2H), 2.56 (d, J=6.3 Hz, 2H), 2.12 (s, 3H). LCMS (m/z): 398.1 (M+1).

Compound 343-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoethoxy)benzamide

Step 1: ethyl 2-(3-carbamoylphenoxy)acetate

To a stirred mixture of 3-hydroxybenzamide (300 mg, 2.19 mmol) in MeCN(5 mL) was added ethyl bromoacetate (545 mg, 3.29 mmol) and K₂CO₃ (907mg, 6.57 mmol). The mixture was stirred at 80° C. for 4 hours. Themixture was filtered and the filtrate concentrated. The residue wasdirectly for the next step. LCMS (m/z): 224.1 (M+1).

Step 2:3-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoethoxy)benzamide

To a stirred mixture of ethyl 2-(3-carbamoylphenoxy)acetate (150 mg,0.673 mmol) in EtOH (1 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (138.6 mg, 0.673mmol). The mixture was stirred at 120° C. for 0.5 hour under mediatedheating. After evaporation of the solvent, the residue was purified byprep-HPLC to afford the desired title product (64 mg, Yield 25%). ¹H NMR(400 MHz, MeOD) δ 7.53 (d, J=8.0 Hz, 1H), 7.48 (s, 1H), 7.39 (dd,J₁=J₂=7.9 Hz, 1H), 7.12-7.00 (m, 5H), 4.60 (s, 2H), 4.04-4.01 (m, 1H),3.74-3.65 (m, 2H), 3.47-3.39 (m, 2H), 2.95-2.87 (m, 2H), 2.85-2.77 (m,2H), 2.58 (d, J=6.0 Hz, 2H). LCMS (m/z): 384.1 (M+1).

Compound 46N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetamide

Step 1: 5-methoxy-1-methyl-1H-benzo[d]imidazole

To a solution of NaH (972 mg, 40.5 mmol) in DMF (20 mL) was added5-methoxy-1H-benzo[d]imidazole (2.0 g, 13.5 mmol) at 27° C. Afterstirring for 5 minutes, MeI (2.3 g, 16.2 mmol) was added and theresulting mixture was stirred for 16 h. The mixture was then dilutedwith water (100 mL) and extracted with ethyl acetate (2×50 mL). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the crude product (1.2 g, 54.5%) as agrown solid. This crude was used in next step without furtherpurification. LCMS (m/z): 163.1 (M+1).

Step 2: 1-methyl-1H-benzo[d]imidazol-5-ol

To a solution of 5-methoxy-1-methyl-1H-benzo[d]imidazole (500 mg, 3.08mmol) in CH₂Cl₂ (6 ml) was added BBr₃ (3.1 g, 12.33 mmol) dropwise at 0°C. After addition, the mixture was stirred for 2 h at 0° C. The mixturewas then quenched by slow addition to ice water (50 mL). The resultingmixture was extracted with CH₂Cl₂ (2×20 mL). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄ and concentrated togive the title compound (100 mg, 21.9%) as a white solid which was usedin next step without further purification. LCMS (m/z): 149.1 (M+1).

Step 3: 2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetic acid

To a solution of NaH (204 mg, 8.49 mmol) in DMF (5 mL) was added1-methyl-1H-benzo[d]imidazol-5-ol (420 mg, 2.83 mmol) at 28° C. Afterbeing stirred for 5 minutes, ethyl 2-bromoacetate (568 mg, 3.4 mmol) wasadded and the resulting mixture stirred for a further 16 h under thereaction was complete by TLC. The mixture was treated with water (50 mL)and extracted with ethyl acetate (2×20 mL). The water layer was treatedwith 2N HCl until pH 3 and extracted with ethyl acetate (2×20 mL). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the product (160 mg, 24.1%) as whitesolid which was used in next step without further purification. LCMS(m/z): 207.1 (M+1).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetamide

To a solution of 2-((1-methyl-1H-benzo[d]imidazol-5-yl)oxy)acetic acid(160 mg, 0.776 mmol) in DMF (4 ml) was added TEA (336 mg, 3.33 mmol),HOBt (157 mg, 1.164 mmol), EDCI (223 mg, 1.164 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (192 mg, 0.931mmol) at 29° C. The reaction mixture was stirred for 16 h at 29° C.until TLC showed the reaction to be complete. After evaporation of thesolvent, the mixture was purified by preparative HPLC to give the titlecompound (13.1 mg, 4.2%) as colorless oil. ¹H NMR (400 MHz, MeOD) δ 8.05(s, 1H), 7.45 (d, J=8.8 Hz, 1H), 7.21 (d, J=2.0 Hz, 1H), 7.09-6.97 (m,5H), 4.59 (s, 2H), 4.03-3.97 (m, 1H), 3.84 (s, 3H), 3.65 (dd, J1=14.8Hz, J2=30.4 Hz, 2H), 3.47-3.42 (m, 2H), 2.88 (t, J=5.6 Hz, 2H), 2.72 (t,J=5.6 Hz, 2H), 2.53-2.51 (m, 2H). LCMS (m/z): 395.2 (M+1).

Compound 37N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-(methylsulfonyl)phenoxy)acetamide

Step 1: ethyl 2-(2-(methylsulfonyl)phenoxy)acetate

To a solution of 2-(methylsulfonyl)phenol (200 mg, 1.16 mmol) in CH₃CN(10 mL) was added ethyl 2-bromo-2-methylpropanoate (232 mg, 1.39 mmol)and K₂CO₃ (690 mg, 5 mmol) at 25° C. The mixture was refluxed for 16 hand then quenched by addition of water (100 mL). The resulting mixturewas extracted with ethyl acetate (2×50 mL). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄ and concentrated togive the title compound which was used in next step without furtherpurification.

Step 2: 2-(2-(methylsulfonyl)phenoxy)acetic acid

To a solution of ethyl 2-(2-(methylsulfonyl)phenoxy)acetate (750 mg, 2.9mmol) in EtOH (15 ml) was added 10% NaOH aqueous solution (15 mL) at 26°C. The mixture was stirred for 30 min and then concentrated and theresidue diluted with water (20 mL) and washed with ethyl acetate (2×20mL). The aqueous layer was then acidified with 2N HCL until pH 3 andthen extracted with ethyl acetate (2×20 ml). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄ and concentrated togive the title compound which was used in next step without furtherpurification.

Step 3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-(methylsulfonyl)phenoxy)acetamide

A mixture of compound 2-(4-methoxyphenoxy)acetic acid (100 mg, 0.43mmol), 1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (89 mg,0.43 mmol), BOP-Cl (171 mg, 0.67 mmol) and DIPEA (1 mL) in DCM (10 mL)was stirred at room temperature for 4 h. The solvent was removed byconcentration and the crude product was purified by pre-HPLC to give thetitle compound (12 mg, 6.7%). ¹H NMR (500 MHz, MeOD): δ 7.96 (dd, J=1.6,8.0 Hz, 1H), 7.75 (t, J=7.2 Hz, 1H), 7.29-7.24 (m, 2H), 7.13-7.02 (m,4H), 4.82 (s, 2H), 4.06-4.03 (m, 1H), 3.72 (d, J=2.4 Hz, 2H), 3.53 (dd,J=7.2, 13.2 Hz, 1H), 3.37-3.33 (m, 1H), 3.29 (s, 3H), 2.93-2.90 (m, 2H),2.86-2.83 (m, 2H), 2.62-2.6 (m, 2H) ppm; ESI-MS (m/z): 419.1 [M+1]⁺.

Compound 39N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

Step 1: ethyl 2-(quinolin-8-yloxy)propanoate

To a solution of compound NaH (100 mg, 4.14 mmol) in DMF (3 mL) wasadded quinolin-8-ol (200 mg, 1.38 mmol) at 26° C. After stirred for 5minutes, ethyl 2-bromopropanoate (300 mg, 1.65 mmol) was added and thereaction mixture stirred for 16 h at 26° C. The mixture was then dilutedwith water (20 mL) and extracted with ethyl acetate (3×10 mL). Thecombined organic layers were washed with brine (20 mL), dried overNa₂SO₄ and concentrated to give ethyl 2-(quinolin-8-yloxy)propanoate(200 mg, 59.2%) as colorless oil which was used in next step withoutfurther purification. (304 mg, Yield 90%).

Step 2: 2-(quinolin-8-yloxy)propanoic acid

To a solution of ethyl 2-(quinolin-8-yloxy)propanoate (100 mg, 0.4 mmol)in EtOH (1 ml) was added a solution of NaOH (24 mg, 0.6 mmol) in H₂O(0.5 ml) at 27° C. The mixture was stirred for 30 min at 27° C. Themixture was then concentrated and the residue treated with water (5 mL)and extracted with ethyl acetate (2×5 mL). The water layer was thentreated with 2N HCl until pH 3 before being extracted with ethyl acetate(2×5 ml). The organic layer was washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the title product (70 mg, 80.5%) as awhite solid which was used in next step without further purification.

Step 3:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

To a solution of 2-(quinolin-8-yloxy)propanoic acid (60 mg, 0.276 mmol)in DMF (4 ml) was added TEA (84 mg, 1.1 mmol), HOBt (60 mg, 0.41 mmol),EDCI (79.8 mg, 0.41 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (72 mg, 0.331mmol) at 28° C. The reaction mixture was stirred for 16 h until TLCshowed the reaction was completed. After evaporation of the solvent, theresidue was purified by HPLC separation to give the TFA salt ofN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide(23 mg, 20.5%) as a white solid. ¹H NMR (400 MHz, MeOD) δ 9.15 (d, J=4.0Hz, 1H), 9.04-9.00 (m, 1H), 8.06-8.02 (m, 1H), 7.89-7.80 (m, 2H), 7.57(d, J=7.6 Hz, 1H), 7.33-7.19 (m, 4H), 5.29-5.27 (m, 1H), 4.50 (br.s,2H), 4.30-4.27 (m, 1H), 3.69 (br.s, 2H), 3.45-3.42 (m, 2H), 3.26-3.20(m, 4H), 2.76 (d, J=9.6 Hz, 3H). LCMS (m/z): 406.2 (M+1).

Compound 43N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-(quinolin-8-yloxy)propanamide

Step 1: ethyl 2-methyl-2-(quinolin-8-yloxy)propanoate

To a solution of compound NaH (248 mg, 10.2 mmol) in DMF (10 mL) wasadded quinolin-8-ol (500 mg, 3.44 mmol) at 28° C. After stirring for 5minutes, ethyl 2-bromo-2-methylpropanoate (806 mg, 4.13 mmol) was addedand the reaction mixture was stirred for an additional 16 h at 28° C.until the reaction was complete by TLC. The mixture was then dilutedwith water (50 mL) and extracted with ethyl acetate (3×20 mL). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the crude product (400 mg, 44.8%) ascolorless oil which was used in next step without further purification.

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-methyl-2-(quinolin-8-yloxy)propanamide

To a solution of ethyl 2-methyl-2-(quinolin-8-yloxy)propanoate (120 mg,0.46 mmol) in EtOH (0.5 ml) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (0.46 mmol) at29° C. The mixture was stirred for 1 hour at 120° C. under microwaveheating. The solvent was removed and the residue purified by prep-HPLCto afford the title compound (19.5 mg, Yield 10.1%). ¹H NMR (400 MHz,MeOD): δ 8.95-8.93 (m, 1H), 8.34 (dd, J=8.4, 1.6 Hz, 1H), 7.69 (d, J=8Hz, 1H), 7.56-7.52 (m, 2H), 7.39 (d, J=7.6 Hz, 1H), 7.11-7.02 (m, 3H),6.94 (d, J=6.4 Hz, 1H), 4.13-4.10 (m, 1H), 3.70-3.53 (m, 2H), 3.48-3.43(m, 2H), 2.91-2.81 (m, 4H), 2.79-2.64 (m, 2H), 1.57 (s, 6H) ppm; ESI-MS(m/z): 420.3 [M+1]⁺.

Compound 44(S)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)acetamide

To a stirred mixture of ethyl 2-(quinolin-8-yloxy)acetate (250 mg, 1.08mmol) in EtOH (2 mL) was added(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (222 mg, 1.08mmol). The mixture was stirred at 120° C. for 0.5 hour under microwaveheating. After evaporation of the solvent, the residue was purifiedfirst by prep-TLC and then prep-SFC to afford (140 mg, Yield 36%). ¹HNMR (400 MHz, MeOD) δ=8.91 (d, J=4.3 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H),7.69-7.54 (m, 3H), 7.30 (d, J=7.3 Hz, 1H), 7.12-6.96 (m, 4H), 4.78 (s,2H), 4.18-4.07 (m, 1H), 3.71 (s, 2H), 3.60-3.49 (m, 1H), 3.48-3.40 (m,1H), 2.89 (d, J=5.8 Hz, 2H), 2.84 (d, J=4.8 Hz, 2H), 2.69-2.55 (m, 2H).LCMS (m/z): 392.1 (M+1).

Compound 45(R)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)acetamide

To a stirred mixture of ethyl 2-(quinolin-8-yloxy)acetate (250 mg, 1.08mmol) in EtOH (2 mL) was added(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (222 mg, 1.08mmol). The mixture was stirred at 120° C. for 0.5 hour under microwaveheating. After evaporation of the solvent, the residue was purifiedfirst by prep-TLC and then by prep-SFC to afford (160 mg Yield 40%). ¹HNMR (400 MHz, MeOD) δ=8.791 (d, J=4.3 Hz, 1H), 8.30 (d, J=8.3 Hz, 1H),7.51-7.46 (m, 3H), 7.17 (d, J=7.3 Hz, 1H), 6.94-6.85 (m, 4H), 4.65 (s,2H), 4.00-3.99 (m, 1H), 3.59 (s, 2H), 3.44-3.32 (m, 2H), 2.77-2.71 (m,4H), 2.53-2.51 (m, 2H). LCMS (m/z): 392.1 (M+1).

Compound 48N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-morpholinoquinolin-8-yl)oxy)acetamide

Step 1: 2-chloroquinolin-8-ol

To a stirred mixture of quinoline-2,8-diol (1 g, 6.21 mmol) was addedPOCl₃ (10 mL) and the mixture stirred at 100° C. for 1 hour beforecooling. The mixture was then poured into ice-water (100 mL) slowly andfiltered. The collected solid was dried and used in next step withoutfurther purification. (780 mg, Yield 70%). ¹H NMR (400 MHz, DMSO-d₆) δ8.39 (d, J=8.8 Hz, 1H), 7.57 (d, J=8.8 Hz, 1H), 7.48-7.43 (m, 2H), 7.17(dd, J₁=8.8 Hz, J₁=1.6 Hz, 1H).

Step 2: 2-morpholinoquinolin-8-ol

To a stirred mixture of 2-chloroquinolin-8-ol (1.7 g crude, 9.5 mmol)was added morpholine (5 mL). The mixture was heated at reflux for 16hours. After cooling, the mixture was diluted with water (40 mL) andextracted with ethyl acetate (3×30 mL). The combined organic layers werewashed with brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated.The residue was used directly for the next step. (1.6 g, Yield 80%).LCMS (m/z): 231.1 (M+1).

Step 3: ethyl 2-((2-morpholinoquinolin-8-yl)oxy)acetate

To a stirred mixture of 2-morpholinoquinolin-8-ol (200 mg, 0.87 mmol) inMeCN (5 mL) was added ethyl bromoacetate (216 mg, 1.31 mmol) and K₂CO₃(360 mg, 2.61 mmol). The mixture was stirred at 80° C. for 4 hours.After filtration, the filtrate was concentrated to give crude productwhich was used directly for the next step (250 mg, Yield 90%).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-morpholinoquinolin-8-yl)oxy)acetamide

To a stirred mixture of ethyl 2-((2-morpholinoquinolin-8-yl)oxy)acetate(100 mg, 0.316 mmol) in EtOH (2 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (65 mg, 0.316mmol). The mixture was stirred at 120° C. for 0.5 hour under microwaveconditions then after evaporation of solvent, the reaction mixture waspurified by prep-HPLC to afford the title product (14 mg, Yield 10%). ¹HNMR (400 MHz, MeOD) δ=8.04 (d, J=9.3 Hz, 1H), 7.43-7.36 (m, 1H),7.23-7.16 (m, 3H), 7.10-7.00 (m, 3H), 6.97 (d, J=4.8 Hz, 1H), 4.77 (s,2H), 3.96 (t, J=6.3 Hz, 1H), 3.87-3.83 (m, 4H), 3.77-3.72 (m, 4H),3.63-3.53 (m, 2H), 3.52-3.45 (m, 1H), 3.40 (d, J=6.3 Hz, 1H), 2.83 (d,J=5.8 Hz, 2H), 2.75-2.67 (m, 2H), 2.56-2.37 (m, 2H). LCMS (m/z): 477.2(M+1).

Compound 49N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-morpholinophenoxy)acetamide

Step 1: 4-(2-methoxyphenyl)morpholine

To a solution of 1-iodo-2-methoxybenzene (1 g, 4.28 mmol) in dioxane (10mL) was added morpholine (446.8 mg, 5.12 mmol), Pd₂(dba)₃ (100 mg, 0.1mmol), Xantphos (200 mg, 0.3 mmol) and t-BuONa (671 mg, 6.0 mmol). Undera N₂ atmosphere the reaction mixture was heated at reflux temperaturefor 16 h. The solvent was then removed and the residue dissolved inethyl acetate and washed with water. The separated organic layer wasconcentrated to give the crude product which was used in next stepwithout further purification (578 mg Yield 70%). LCMS (m/z): 194.1(M+1).

Step 2: 2-morpholinophenol

To a solution of 4-(2-methoxyphenyl)morpholine (200 mg, 1.02 mmol) inCH₂Cl₂ (20 mL) was added BBr₃ (1 mL) at 0° C. The mixture was stirredfor 2 h at 0° C. The mixture was then added drop wise to ice-water (50mL) and the mixture treated with CH₂Cl₂ (2×20 mL). The combined organiclayers were washed with brine (30 mL), dried over Na₂SO₄ andconcentrated to give the yellow solid which was used in next stepwithout further purification (165 mg Yield 80%). LCMS (m/z): 180.1(M+1).

Step 3: ethyl 2-(2-morpholinophenoxy)acetate

A mixture of 2-morpholinophenol (100 mg, 0.56 mmol) and ethyl2-bromoacetate (200 mg, 0.672 mmol) in CH₃CN (10 mL) was added K₂CO₃(772.8 mg, 5.6 mmol). The reaction mixture was stirred at 80° C. for 4h. The solid was removed by filtration and the filtrate concentrated togive a crude material, which was used in the next step without furtherpurification. (130 mg, Yield 90%).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-morpholinophenoxy)acetamide

A mixture of ethyl 2-(2-morpholinophenoxy)acetate (53 mg, 0.2 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (41 mg, 0.2 mmol)in EtOH (1 mL) was stirred at 120° C. over microwave for 30 min. Thesolvent was removed by concentration and the crude product was purifiedby prep-HPLC separation to afford product the desired title compound(8.0 mg, Yield 10%). ¹H NMR (400 MHz, MeOD): 7.09-6.98 (m, 8H), 4.64 (s,2H), 3.95 (br.s, 1H), 3.89-3.87 (m, 4H), 3.63-3.46 (m, 2H), 3.33-3.30(m, 1H), 3.07-3.03 (m, 4H), 2.88 (dd, J=6.0 Hz, 2H), 2.77 (dd, J=6.0 Hz,2H), 2.49 (d, J=6.0 Hz, 2H). LCMS (m/z): 426.2 (M+1).

Compound 502-(2-(1H-pyrazol-3-yl)phenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

Step 1: ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate

To a solution of 2-(1H-pyrazol-3-yl)phenol (500 mg, 3.125 mmol), K₂CO₃(517.5 mg, 3.75 mmol) and ethyl 2-bromoacetate (417.5 mg, 2.5 mmol) inMeCN (20 mL). The mixture was stirred at room temperature for 2 h, atwhich time TLC showed the completion of the reaction. The mixture wasdiluted with water and extracted with EtOAc. The combined organic layerswere washed with brine, dried over Na₂SO₄ and concentrated to give thecrude product which was used in next step without further purification.

Step 2:2-(2-(1H-pyrazol-3-yl)phenoxy)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)acetamide

To a solution of ethyl 2-(2-(1H-pyrazol-3-yl)phenoxy)acetate (100 mg,0.41 mmol) in EtOH (10 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (84 mg, 0.41mmol). The mixture was stirred at 120° C. under microwave heating for 2h. The reaction mixture was diluted with ethyl acetate (30 mL) andwashed with water (10 mL), dried over Na₂SO₄ and concentrated to givethe crude product. The residue was purified by prep-HPLC to afford thedesired title compound (85 mg, 44%). ¹H NMR (400 MHz, MeOD): δ 7.71-7.69(m, 2H), 7.34-7.33 (m, 1H), 7.10-7.01 (m, 6H), 6.74 (d, J=2 Hz, 1H),4.67 (s, 1H), 4.04-4.02 (m, 1H), 3.67 (s, 2H), 3.50-3.49 (m, 1H),3.37-3.33 (m, 1H), 2.89-2.87 (m, 2H), 2.81-2.78 (m, 2H), 2.58-2.57 (m,2H) ppm; ESI-MS (m/z): 469.3 [M+1]⁺.

Compound 54N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetamide

Step 1: ethyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate

A mixture of ethyl 2-(3-bromophenyl)acetate (1.0 g, 4.1 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.34 g, 5.3mmol), KOAc (862 mg, 8.8 mmol) and Pd(pddf)Cl₂ (50 mg) in dioxane (15mL) was stirred at 120° C. for 16 h under N₂. The reaction mixture wasconcentrated and the residue dissolved in water then extracted withEtOAc. The organic layer was concentrated, and the residue purified bycolumn chromatography to give the product which was used directly in thenext step.

Step 2: ethyl 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetate

A mixture of ethyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate (500mg, 1.72 mmol), 5-bromo-1-methyl-1H-pyrazole (252 mg, 1.57 mmol), K₂CO₃(651 mg, 4.71 mmol) and Pd(dppf)Cl₂ (20 mg) in a solution of dioxane (10mL) and H₂O (2.5 mL) was stirred at 120° C. for 30 min under microwave.The catalyst was filtered through a pad of celite and the filtrateconcentrated. The residue was purified by column chromatography to givethe desired product (270 mg, Yield 70%) and used directly in the nextstep. LCMS (m/z): 245.1 (M+1).

Step 3: 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetic acid

To a solution of ethyl 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetate(300 mg, 1.2 mmol) in MeOH (6 mL) was added aqueous NaOH (1.5 mL, 40 W%). The mixture was stirred at room temperature for 2 h. The reactionmixture was concentrated and the residue dissolved in water and adjustedpH to 5-6 with 2N of HCl. The solution was then extracted with EtOAc andthe combined organic layers were concentrated to give the crude productwhich was used directly in the next step. LCMS (m/z): 231.1 (M+1).

Step 4:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetamide

To a solution of 2-(3-(1-methyl-1H-pyrazol-5-yl)phenyl)acetic acid (150mg, 0.69 mmol) in DCM (6 mL) was added EDCI (265 mg, 1.38 mmol), HOBt(186 mg, 1.38 mmol), Et₃N (209 mg, 2.07 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (142 mg, 0.69mmol). The mixture was stirred at room temperature for 16 h then dilutedwith water (10 mL) and extracted with DCM (10 mL×3). The combinedorganic layers were concentrated. The residue was purified by prep-HPLCto give the product as a colorless oil (60 mg, Yield 21%). ¹H NMR (400MHz, MeOD): 7.47 (s, 1H), 7.43-7.33 (m, 4H), 7.08-7.04 (m, 3H),6.96-6.94 (m, 1H), 6.35 (s, 1H), 3.96-3.91 (m, 1H), 3.83 (s, 3H),3.60-3.59 (m, 4H), 3.38-3.20 (m, 2H), 2.84 (t, J=6.0 Hz, 2H), 2.72 (t,J=6.0 Hz, 2H), 2.49 (d, J=6.4 Hz, 2H). LCMS (m/z): 405.2 (M+1).

Compound 60N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yl)acetamide

A solution of 2-(quinolin-8-yl)acetic acid (187 mg, 1 mmol), HATU (387.6mg, 1.02 mmol) and TEA (196.1 mg, 1.94 mmol) in DCM (10 mL) was stirredat room temperature for 10 min.1-Amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (210 mg, 1.0mmol) was then added, and the solution stirred at for another 3 h, atwhich point LCMS indicated completion of the reaction. The reactionmixture was diluted with water and extracted with DCM (10 mL×3). Theorganic layers combined and dried over anhydrous Na₂SO₄, filtered andconcentrated. The residue was purified by prep-HPLC to give the desiredcompound (50 mg, Yield 13%). ¹H NMR (400 MHz, MeOD): 8.92 (d, J=2.8 Hz,1H), 8.33 (d, J=8.0 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.73 (d, J=6.8 Hz,1H), 7.55-7.50 (m, 2H), 7.10-6.97 (m, 4H), 4.25 (dd, J₁=10.8 Hz, J₂=14.0Hz, 2H), 3.90 (m, 1H), 3.54-3.51 (m 2H), 3.32-3.25 (m, 2H), 2.82-2.80(m, 2H), 2.67-2.66 (m, 2H), 2.40-2.39 (m, 2H). LCMS (m/z): 376.1 (M+1).

Compound 62N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetamide

Step 1: ethyl 2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetate

To a stirred mixture of 5,6,7,8-tetrahydronaphthalen-1-ol (200 mg, 1.35mmol) in MeCN (5 mL) was added ethyl bromoacetate (269 mg, 1.62 mmol)and K₂CO₃ (372 mg, 2.70 mmol). The mixture was stirred at 80° C. for 4hours. The mixture was filtered, the filtrate concentrated to yield thedesired product which used directly for the next step without furtherpurification (300 mg, Yield 95%).

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetamide

To a stirred mixture of ethyl2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetate (150 mg, 0.641 mmol)in EtOH (2 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg, 0.641mmol). The mixture was stirred at 120° C. for 0.5 hours under microwaveheating. After evaporation of the solvent, the residue was purified byprep-HPLC to afford the desired target productN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((5,6,7,8-tetrahydronaphthalen-1-yl)oxy)acetamide(14 mg, Yield 6%). ¹H NMR (400 MHz, MeOD) δ=7.13-7.00 (m, 5H), 6.74 (d,J=7.8 Hz, 1H), 6.65 (d, J=8.3 Hz, 1H), 4.53 (s, 2H), 4.02 (quin, J=5.9Hz, 1H), 3.72-3.63 (m, 2H), 3.53-3.45 (m, 1H), 3.39 (d, J=6.3 Hz, 1H),2.96-2.86 (m, 2H), 2.83-2.69 (m, 6H), 2.56 (d, J=6.3 Hz, 2H), 1.84-1.74(m, 4H). LCMS (m/z): 395.1 (M+1).

Compound 66N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(naphthalen-1-yloxy)acetamide

Step 1: ethyl 2-(naphthalen-1-yloxy)acetate

To a stirred mixture of naphthalen-1-ol (196 mg, 1.35 mmol) in MeCN (5mL) was added ethyl bromoacetate (269 mg, 1.62 mmol) and K₂CO₃ (372 mg,2.70 mmol). The mixture was stirred at 80° C. for 4 hours. The mixturewas filtered and the filtrate concentrated. The residue was useddirectly for the next step without further purification (300 mg, Yield95%).

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(naphthalen-1-yloxy)acetamide

To a stirred mixture of ethyl 2-(naphthalen-1-yloxy)acetate (150 mg,0.641 mmol) in EtOH (2 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (132 mg, 0.641mmol). The mixture was stirred at 120° C. for 30 minutes under microwavemediated heating. After evaporation of the solvent, the reaction mixturewas purified by prep-HPLC to afford the desired product (64 mg, Yield25%). ¹H NMR (400 MHz, METHANOL-d₄) δ=8.41-8.31 (m, 1H), 7.90-7.80 (m,1H), 7.58-7.46 (m, 3H), 7.44-7.35 (m, 1H), 7.18-6.95 (m, 5H), 6.91 (d,J=7.8 Hz, 1H), 4.76 (s, 2H), 4.05 (quin, J=6.0 Hz, 1H), 3.72-3.59 (m,2H), 3.56-3.48 (m, 1H), 3.41 (dd, J=6.5, 13.6 Hz, 1H), 2.93-2.83 (m,2H), 2.80-2.72 (m, 2H), 2.59-2.52 (m, 2H). LCMS (m/z): 391.2 (M+1).

Compound 71N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(1-(1-methylpiperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide

Step 1: tert-butyl 4-((methylsulfonyl)oxy)piperidine-1-carboxylate

To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (3.0 g,14.9 mmol) in DCM (30 mL) was added triethylamine (4.5 g, 44.8 mmol). Tothis mixture methanesulfonyl chloride (5.1 g, 44.8 mmol) was addeddropwise. After addition, the mixture was stirred at 25° C. for 3 h andthen filtered. The filtrate was washed with aqueous HCl, dried overNa₂SO₄, and concentrated under reduced pressure to give tert-butyl4-((methylsulfonyl)oxy)piperidine-1-carboxylate (3.58 g, yield 86%) as awhite solid. This material was used in the next step without furtherpurification. LCMS (m/z): 280.2 [M+H]⁺

¹H NMR (400 MHz, CDCl₃) δ ppm 4.75-4.82 (m, 1H) 3.60-3.71 (m, 2H)3.21-3.32 (m, 2H) 2.95 (s, 3H) 1.78-1.85 (m, 2H) 1.67-1.78 (m, 2H) 1.35(s, 9H)

Step 2 5-((tert-butyldimethylsilyl)oxy)-1H-indazole

To a solution of 1H-indazol-5-ol (400 mg, 2.98 mmol) in DMF (10 mL) wasadded TBDMSCl (537 mg, 3.58 mmol) and imidazole (405 mg, 5.96 mmol) at0° C. and the resulting mixture was stirred at 25° C. for 16 h. Thereaction was quenched by addition of water and the product extractedwith ethyl acetate. The organic phase was washed with brine, then driedover Na₂SO₄, and concentrated under reduced pressure to give5-((tert-butyldimethylsilyl)oxy)-1H-indazole (500 mg, yield 68%) as abrown solid, which was used in the next step without furtherpurification. LCMS (m/z): 249.1 [M+H]⁺

Step 3 tert-butyl 4-(5-hydroxy-1H-indazol-1-yl)piperidine-1-carboxylate

To a solution of NaH (60% in mineral oil) (43.5 mg, 1.81 mmol) in DMF at0° C. was added 5-((tert-butyldimethylsilyl)oxy)-1H-indazole (300 mg,1.21 mmol), and the mixture was stirred at 0° C. for 15 min. Tert-butyl4-((methylsulfonyl)oxy)piperidine-1-carboxylate (243 mg, 1.21 mmol) wasthen added to the mixture at 0° C. After addition, the mixture wasstirred at 85° C. for 12 h. The mixture was poured into water, and theproduct extracted with ethyl acetate. The organic phase was washed withwater, dried over Na₂SO₄, concentrated under reduced pressure, andpurified by TLC (Pet.Ether:EtOAc=2:1) to give tert-butyl4-(5-hydroxy-1H-indazol-1-yl)piperidine-1-carboxylate (250 mg, yield65%) as a colorless oil. LCMS (m/z): 318.2 [M+H]⁺

Step 4 tert-butyl4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate

To a solution of tert-butyl4-(5-hydroxy-1H-indazol-1-yl)piperidine-1-carboxylate (150 mg, 0.458mmol) in DMF (10 mL) at 0° C. was added NaH (60% in mineral oil) (16.8mg, 0.706 mmol) and ethyl 2-bromoacetate (119 mg, 0.706 mmol). Themixture was stirred at 25° C. for 3 h, poured into water, and theproduct extracted with ethyl acetate. The organic phase was washed withwater, dried over Na₂SO₄, concentrated under reduced pressure andpurified by TLC (Pet.Ether:EtOAc=1:1) to give tert-butyl4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate(120 mg, yield 65%) as a colorless oil. LCMS (m/z): 404.2 [M+H]⁺

Step 5 tert-butyl4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate

To a solution of tert-butyl4-(5-(2-ethoxy-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate(100 mg, 0.248 mmol) in EtOH (2 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (102.2 mg, 0.496mmol) and the mixture was stirred at 120° C. for 2 h in microwave underN₂. The mixture was allowed to cool, concentrated under reducedpressure, purified by TLC (Pet.Ether:EtOAc=1:1) to give tert-butyl4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate(70 mg, yield 50%) as a colorless oil. LCMS (m/z): 564.3 [M+H]⁺

Compound 70N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide

To tert-butyl4-(5-(2-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)amino)-2-oxoethoxy)-1H-indazol-1-yl)piperidine-1-carboxylate(110 mg, 0.195 mmol) was added EtOAc.HCl (10 mL), the solution wasstirred at 25° C. for 2 h, concentrated under reduced pressure, andpurified by prep-HPLC to giveN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide(85 mg, yield 94%) as a colorless oil. ¹H NMR (400 MHz, METHANOL-d₄) δppm 8.47 (br. s., 2H), 8.00 (s, 1H), 7.70 (d, J=8.78 Hz, 1H), 7.29-7.19(m, 3H), 7.18-7.11 (m, 2H), 6.93 (dd, J=8.85, 1.95 Hz, 1H), 5.12 (s,2H), 4.86-4.81 (m, 1H), 4.17 (s, 3H), 3.49-3.35 (m, 4H), 3.32-3.20 (m,4H), 3.13-2.92 (m, 4H), 2.28-2.04 (m, 4H). LCMS (m/z): 464.2 [M+H]⁺

Compound 71N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-O-(1-methylpiperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide

To a solution ofN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(piperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide(50 mg, 0.108 mmol) in MeOH (5 mL) was added triethylamine (1 mL), HCHO(30%)(0.3 mL), and HOAC (0.4 mL). The mixture was stirred at 25° C. for30 min, then NaBH₃CN (0.4 mg) was added, and the mixture was stirred at25° C. for an additional 1 h, concentrated under reduced pressure, andpurified by prep-HPLC to giveN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((1-(1-methylpiperidin-4-yl)-1H-indazol-5-yl)oxy)acetamide(51.1 mg, yield 99%) as a colorless oil. ¹H NMR (400 MHz, METHANOL-d₄) δppm 8.50 (br. s., 2H), 8.00 (s, 1H), 7.70 (d, J=8.78 Hz, 1H), 7.28-7.19(m, 3H), 7.17-7.11 (m, 2H), 6.94 (dd, J=8.85, 1.82 Hz, 1H), 5.12 (s,2H), 4.82 (br. s., 2H), 4.24-4.11 (m, 3H), 3.47-3.35 (m, 3H), 3.32 (br.s., 1H), 3.13-2.94 (m, 4H), 2.87 (s, 3H), 2.32-2.07 (m, 4H). LCMS (m/z):478.3 [M+H]⁺

Compound 73(R)-N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

Step 1: (R)-Methyl 2-(quinolin-8-yloxy)propanoate

To a stirred mixture of quinolin-8-ol (300 mg, 2.07 mmol) in THF (5 mL)was added (S)-methyl 2-hydroxypropanoate (215 mg, 2.07 mmol), PPh₃ (647mg, 2.47 mmol) and DEAD (430 mg, 2.47 mmol). The mixture was stirred at25° C. for 16 hours. Subsequently, 1M HCl was added (10 mL) and thesolution was washed with EtOAc (20 mL×3). The pH of the aqueous solutionwas raised by addition of aqueous NaHCO₃ (10 mL), and then this solutionwas washed with EtOAc (10 mL×3). The combined organic extracts werewashed with brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated.The residue was purified by silica column chromatography to afford theproduct as a colorless oil (300 mg, 62.5% yield). LCMS (m/z): 233.1[M+H]⁺

Step 2:(R)-N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

To a stirred mixture of (R)-methyl 2-(quinolin-8-yloxy)propanoate (100mg, 0.433 mmol) in EtOH (1 mL) was added(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl) propan-2-ol (89.2 mg,0.433 mmol). The mixture was stirred in a sealed tube in a microwaveapparatus at 120° C. for 0.5 hour. After cooling to room temperature thesolvent was evaporated, and the residue was first purified by prep-TLCand then prep-HPLC to afford the title compound (90 mg, yield: 51%). ¹HNMR (400 MHz, METHANOL-d₄) δ ppm 8.89 (br. S., 1H), 8.29-8.42 (m, 1H),7.49-7.65 (m, 3H), 7.28 (d, J=5.52 Hz, 1H), 6.89-7.13 (m, 4H), 5.01-5.13(m, 1H), 4.10-3.84 (m, 1H), 3.45-3.60 (m, 2H), 3.35-3.43 (m, 2H), 2.81(d, J=3.01 Hz, 2H), 2.65 (br. S., 2H), 2.34-2.49 (m, 2H) 1.71 (d, J=6.78Hz, 3H), LCMS (m/z): 406.2 [M+H]⁺

Compound 76(S)-N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

Step 1: (S)-methyl 2-(quinolin-8-yloxy)propanoate

To a stirred mixture of quinolin-8-ol (300 mg, 2.07 mmol) in THF (5 mL)was added (R)-methyl 2-hydroxypropanoate (215 mg, 2.07 mmol), PPh₃ (647mg, 2.47 mmol) and DEAD (430 mg, 2.47 mmol). The mixture was stirred at25° C. for 16 hours. Subsequently, 1M HCl was added (10 mL) and thesolution was washed with EtOAc (20 mL×3). The pH of the aqueous solutionwas raised by addition of aqueous NaHCO₃ (10 mL), and then this solutionwas washed with EtOAc (10 mL×3). The combined organic extracts werewashed with brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated.The residue was directly for the next step. LCMS (m/z): 232.1/233.1[M+H]⁺

Step 2:(S)-N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(quinolin-8-yloxy)propanamide

To a stirred mixture of (S)-methyl 2-(quinolin-8-yloxy)propanoate (100mg, 0.433 mmol) in EtOH (1 mL) was added(S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (89.2 mg,0.433 mmol). The mixture was stirred in a sealed tube in a microwaveapparatus at 120° C. for 0.5 hour. After cooling to room temperature thesolvent was evaporated, and the residue was first purified by prep-TLCand then prep-HPLC to afford the title compound (49 mg, yield: 28%). ¹HNMR (400 MHz, METHANOL-d₄) δ ppm 8.86-8.97 (m, 1H), 8.38 (d, J=7.03 Hz,1H), 7.51-7.67 (m, 3H), 7.30 (d, J=7.28 Hz, 1H), 7.06-7.13 (m, 3H), 6.98(d, J=6.53 Hz, 1H), 5.07 (q, J=6.53 Hz, 1H), 4.10-3.88 (m, 1H),3.65-3.76 (m, 2H), 3.43-3.51 (m, 1H), 3.35 (br. s., 1H), 2.86 (dd,J=16.06, 3.76 Hz, 4H), 2.62 (d, J=6.02 Hz, 2H), 1.71 (d, J=6.53 Hz, 3H).LCMS (m/z): 406.2 [M+H]⁺

Compound 80N-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetamide

Step 1 2-Aminobenzene-1,3-diol

A solution of 2-nitrobenzene-1,3-diol (5.00 g, 32.2 mmol) in MeOH (100mL) was stirred under H₂ atmosphere (balloon) in the presence of 10%Pd/C (200 mg) for 16 h at room temperature. The reaction mixture wasfiltered and the filtrate was concentrated to render a residuecharacterized as 2-aminobenzene-1,3-diol (3.0 g, 95% yield), used assuch for the next reaction step.

Step 2 5-Hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one

A stirred solution of 2-aminobenzene-1,3-diol (2.0 g, 16.0 mmol) and TEA(1.94 g, 19.2 mmol) in anhydrous DMF (30 mL) was treated with2-chloroacetyl chloride (1.81 g, 16.0 mmol) and stirring continued for16 h at room temperature, then K₂CO₃ (2.65 g, 19.2 mmol) was added andthe mixture further stirred for 16 h at the same temperature. Thereaction mixture was diluted with DCM (100 mL), washed twice with waterand then with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated. The resulting residue was purified by chromatographiccolumn of silicagel to give desired product (1.7 g, 64% yield) LCMS(m/z): 166.1 [M+1]⁺.

Step 3 Ethyl2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate

A stirred mixture of 5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one (100 mg,0.604 mmol) and K₂CO₃ (167 mg, 1.21 mmol) in anhydrous DMF (5 mL) wastreated with ethyl 2-bromoacetate (121 mg, 0.727 mmol) and stirringcontinued at room temperature for 16 h. To this solution of crude ethyl2-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate was addedK₂CO₃ (39.6 mg, 0.287 mmol) followed by MeI (40.7 mg, 0.287 mmol). Afterbeing stirred at room temperature for 16 h, the reaction mixture waspartitioned between water (50 mL) and DCM (100 mL). The organic layerwas washed by water followed by and brine, dried over anhydrous Na₂SO₄,filtered and concentrated and the resulting residue was purified bypreparative TLC to give desired product (43 mg, 56% yield). ¹H NMR (400MHz, CDCl₃): 6.99 (t, J=8.3 Hz, 1H), 6.74 (d, J=7.5 Hz, 1H), 6.55 (d,J=8.3 Hz, 1H), 4.69 (s, 2H), 4.51 (s, 2H), 4.30 (q, J=7.2 Hz, 2H), 3.56(s, 3H), 1.34-1.33 (m, 1H), 1.33 (t, J=7.2 Hz, 3H)

Step 4N-(3-(3,4-Dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetamide

A reaction vessel containing a mixture of ethyl2-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-5-yloxy)acetate(43.0 mg, 0.162 mmol),1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (33.0 mg, 0.163mmol) and EtOH (0.5 mL) was placed in a microwave reactor and themixture irradiated at external temperature of 120° C. for 1 h. Thereaction mixture was purified in two steps by preparative TLC followedby preparative HPLC to render the title product (19.2 mg, 19% yield)

¹H NMR (400 MHz, METHANOL) δ ppm: 8.44 (br. s., 1H), 7.31-7.19 (m, 3H),7.16 (d, J=6.5 Hz, 1H), 7.05 (t, J=1.0 Hz, 1H), 6.76 (dd, J=3.0, 8.3 Hz,2H), 4.76-4.63 (m, 2H), 4.55-4.43 (m, 2H), 4.33-4.17 (m, 3H), 3.50 (s,3H), 3.46-3.36 (m, 4H), 3.17-3.00 (m, 4H). LCMS (m/z): 426.2 [M+H]⁺.

Compound 98(R)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-methoxyquinolin-8-yl)oxy)acetamide

Step 1: 8-(benzyloxy)quinolin-2-ol

To a solution of quinoline-2,8-diol (5.0 g, 31.1 mmol) in i-PrOH (50 mL)was added BnBr (5.31 g, 31.1 mmol) and DBU (2.02 g, 5.32 mmol). Themixture was stirred at 80° C. for 16 h. The mixture was evaporated andto the residue was added DCM (100 mL), and this solution was washed with0.5 N NaOH (50 mL), 10% HCl (50 mL), and H₂O (50 mL). The organic layerwas evaporated to give the desired compound (6.6 g, yield 85%). ¹HNMR(CDCl₃, 400 MHz) δ: 9.16 (br. s., 1H), 7.74 (d, J=9.8 Hz, 1H), 7.49-7.36(m, 5H), 7.20-7.10 (m, 2H), 7.09-7.03 (m, 1H), 6.67 (d, J=9.5 Hz, 1H),5.19 (s, 2H).

Step 2: 8-(benzyloxy)-2-chloroquinoline

8-(benzyloxy)quinolin-2-ol (6.6 g, 26.3 mmol) was dissolved in POCl₃ (50mL). The mixture was stirred at 90° C. for 16 h. The POCl₃ wasevaporated and to the residue was added EtOAc (100 mL) and the solutionwas washed with a.q. NaHCO₃ (80 mL) and H₂O (80 mL). The EtOAc wasremoved under vacuum to give the desired compound (6.0 g, yield 85%).LCMS (m/z): 270.1 [M+H]⁺

Step 3: 8-(benzyloxy)-2-methoxyquinoline

To a solution of MeONa (400 mg, 7.43 mmol) in MeOH (20 mL) was added8-(benzyloxy)-2-chloroquinoline (2.0 g, 7.43 mmol). The mixture wasstirred at 70° C. for 16 h. To the mixture was added H₂O (20 mL) and theproduct extracted with toluene (30 mL×3). The combined organic layerswere dried with Na₂SO₄ and evaporated to give the desired compound (1.5g, yield 79%). LCMS (m/z): 266.1 [M+H]⁺

Step 4: 2-methoxyquinolin-8-ol

To a solution of 8-(benzyloxy)-2-methoxyquinoline (2.2 g, 8.3 mmol) inEtOH (40 mL) was added Pd/C (230 mg). The mixture was stirred at 25° C.for 16 h under an atmosphere of H₂. The mixture was filtered, and thefiltrate was evaporated to give the desired compound (1.2 g, 83%). ¹HNMR(CDCl₃, 400 MHz) δ: 7.91 (d, J=8.8 Hz, 1H), 7.52 (br. s., 1H), 7.23-7.15(m, 2H), 7.07 (dd, J=1.4, 7.2 Hz, 1H), 6.85 (d, J=8.8 Hz, 1H), 3.99 (s,3H).

Step 5: ethyl 2-((2-methoxyquinolin-8-yl)oxy)acetate

To a solution of 2-methoxyquinolin-8-ol (500 mg, 2.86 mmol) in MeCN (10mL) was added ethyl 2-bromoacetate (501 mg, 3.0 mmol) and K₂CO₃ (789 mg,5.72 mmol). The mixture was stirred at 80° C. for 5 h. The mixture wasfiltered and the filtrate evaporated to give the desired compound (700mg, yield 94%). LCMS (m/z): 262.1 [M+H]⁺

Step 6:(R)—N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-methoxyquinolin-8-yl)oxy)acetamide

Ethyl 2-((2-methoxyquinolin-8-yl)oxy)acetate (100 mg, 0.383 mmol),(R)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (79.0 mg,0.383 mmol) and EtOH (1 mL) were combined in a sealed tube. The mixturewas stirred at 120° C. for 30 min in a microwave. The EtOH wasevaporated and the residue was purified by pre-HPLC to give the desiredproduct (77 mg, yield 48%). ¹HNMR (MeOD-d₄, 400 MHz) δ: 8.36 (br. s.,1H), 8.16 (d, J=8.8 Hz, 1H), 7.51 (d, J=7.5 Hz, 1H), 7.36 (t, J=7.9 Hz,1H), 7.28-7.15 (m, 4H), 7.12 (d, J=6.8 Hz, 1H), 7.03 (d, J=8.8 Hz, 1H),4.82 (s, 2H), 4.23-4.16 (m, 3H), 4.10 (s, 3H), 3.54-3.48 (m, 1H),3.47-3.40 (m, 1H), 3.38-3.33 (m, 2H), 3.13-2.98 (m, 4H). LCMS (m/z):422.2 [M+H]⁺

Compound 102N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide

Step 1: 1,2,3,4-tetrahydroisoquinolin-5-ol

A mixture of isoquinolin-5-ol (4 g, 27.6 mmol) and PtO₂ (1.3 g) in HOAc(50 mL) was stirred under H₂ (45 Psi) at room temperature overnight. Themixture was filtered and the filtrate was concentrated under vacuum togive the crude product (3.2 g, 80%) which was used in the next stepwithout purification. LCMS (m/z): 150.1 [M+H]⁺.

Step 2: tert-butyl 3,4-dihydro-5-hydroxyisoquinoline-2(1H)-carboxylate

A mixture of 1,2,3,4-tetrahydroisoquinolin-5-ol (2.06 g, 13.8 mmol) andNa₂CO₃ (2.93 g, 27.6 mmol) in DMF was cooled with an ice-water bath.Then (Boc)₂O (3.61 g, 16.6 mmol) was added in three portions. Thesolution was then stirred at room temperature overnight. The mixture wasthen filtered and the filtrate was concentrated under vacuum to give thecrude product (3.1 g, 91%) which was used directly in the next step.LCMS (m/z): 250.2 [M+H]⁺.

Step 3: tert-butyl5-((ethoxycarbonyl)methoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate

To a solution of tert-butyl3,4-dihydro-5-hydroxyisoquinoline-2(1H)-carboxylate (750 mg, 3.01 mmol)and K₂CO₃ (498 mg, 3.61 mmol) in MeCN was added ethyl 2-bromoacetate(603 mg, 3.61 mmol). The mixture was stirred at room temperatureovernight, the mixture was then filtered and the filtrate wasconcentrated under vacuum to give the desired product (900 mg, 90%).LCMS (m/z): 336.2 [M+H]⁺.

Step 4: ethyl 2-((1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate

To a solution of tert-butyl5-(2-ethoxy-2-oxoethoxy)-3,4-dihydroisoquinoline-2(1H)-carboxylate (400mg, 1.19 mmol) in ethyl acetate (10 mL), cooled in an ice-water bath,was added (10 mL, 1N) drop wise. The mixture was stirred at 25° C. for16 h and then concentrated under vacuum to give the crude product (275mg, 98%) which was used to the next step without further purification.

Step 5: ethyl2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate

To a solution of ethyl2-((1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate (300 mg, 1.28 mmol)and Et₃N (387 mg, 3.83 mmol) in DCM (25 mL) cooled in an ice-water bathwas added MsCl (176.6 mg, 1.54 mmol) drop wise. The mixture was stirredat 25° C. for 16 h and then quenched with aq.NH₄Cl. The mixture wasextracted with ethyl acetate and the combined organic layers wereconcentrated under vacuum to give the crude product (312 mg, 78%) whichwas used to the next step without purification.

Step 6:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide

A mixture of ethyl2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetate(140 mg, 0.447 mmol) and1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (184 mg, 0.89mmol) in EtOH (0.2 mL) was stirred at 120° C. for 30 min under microwaveconditions. The mixture was diluted with MeOH and purified by prep-HPLCto affordN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-((2-(methylsulfonyl)-1,2,3,4-tetrahydroisoquinolin-5-yl)oxy)acetamide(53.5 mg, 25%). ¹H NMR (400 MHz, MeOD) δ 0.84 (s, 1H), 7.29-7.15 (m,5H), 6.80 (d, J=8.2 Hz, 1H), 6.83 (d, J=7.7 Hz, 1H), 4.61 (s, 2H), 4.41(s, 2H), 4.35-4.17 (m, 3H), 3.54 (t, J=6.1 Hz, 2H), 3.48-3.38 (m, 4H),3.17-3.04 (m, 4H), 2.98 (t, J=6.0 Hz, 2H), 2.92 (s, 3H). LCMS (m/z):474.2 [M+H]⁺.

Compound 152N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(pyridin-3-ylmethoxy)acetamide

Step 1: Ethyl 2-(pyridin-3-ylmethoxy)acetate

To a solution of NaH (330 mg, 13.7 mmol) in DMF (10 mL) was addedpyridin-3-ylmethanol (500 mg, 4.6 mmol) and the solution was stirred at27° C. for 20 minutes. Ethyl 2-bromoacetate (921.8 mg, 5.52 mmol) wasthen added and the reaction mixture stirred at 27° C. for further 16 h.Once the reaction was complete by TLC analysis, the mixture was quenchedby addition of water (50 mL) and extracted with ethyl acetate (3×20 mL).The combined organic layers were washed with brine (30 mL), dried overNa₂SO₄ and concentrated to give the Ethyl 2-(pyridin-3-ylmethoxy)acetate(600 mg, 66.8%) as colorless oil which was used in next step withoutfurther purification. LCMS: 196.1 [M+H]⁺.

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(pyridin-3-ylmethoxy)acetamide

To a solution of Ethyl 2-(pyridin-3-ylmethoxy)acetate (100 mg, 0.51mmol) in EtOH (0.5 ml) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (105 mg, 0.51mmol) at 28° C. The mixture was stirred at 120° C. under microwaveheating for 1 hour until the reaction was completed by TLC. Afterevaporation of the solvent, the residue was purified by HPLC separationto give the title compound (30 mg, yield: 16.5%) as a white solid. LCMS:356.2 [M+H]⁺. ¹H NMR (MeOD, 400 MHz) δ 8.53 (s, 1H), 8.50 (d, J=4.8 Hz,1H), 7.85 (d, J=8.0 Hz, 1H), 7.462 (dd, J₁=4.8 Hz, J₂=8.0 Hz, 1H),7.19-7.03 (m, 4H), 4.58 (s, 2H), 4.02 (br. s, 3H), 3.72 (s, 2H),3.42-3.36 (m, 2H), 2.93-2.83 (m, 4H), 2.62-2.61 (m, 2H).

Compound 175 Step 1:(S)-3-(1H-benzo[d]imidazol-2-yl)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)propanamide

To a solution of 3-(1H-benzo[d]imidazol-2-yl)propanoic acid (300 mg,1.579 mmol) in DCM (10 mL) was added HATU (722 mg, 1.895 mmol) and TEA(478 mg, 4.737 mmol). After stirring for 30 min at room temperature,(S)-1-amino-3-(3,4-dihydro isoquinolin-2(1H)-yl)propan-2-ol (488 mg,2.368 mmol) was added and the resulting mixture then stirred at roomtemperature for 16 h. After completion of the reaction the solvent wasevaporated at reduced pressure and the residue purified by preparativeHPLC to give the title compound (159.1 mg, 26%). ¹H NMR (400 MHz,METHANOL-d₄) δ=7.51 (br. s., 2H), 7.22-7.16 (m, 2H), 7.13-7.06 (m, 3H),7.02-6.97 (m, 1H), 3.98-3.90 (m, 1H), 3.66-3.55 (m, 2H), 3.37 (dd,J=4.9, 13.7 Hz, 1H), 3.25-3.17 (m, 3H), 2.89-2.83 (m, 2H), 2.79 (t,J=7.5 Hz, 2H), 2.74-2.69 (m, 2H), 2.50-2.45 (m, 2H). LCMS (m/z): 379.1[M+H]⁺.

Compound 192 Step 1:(R)-3-(1H-benzo[d]imidazol-2-yl)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)propanamide

To a solution of 3-(1H-benzo[d]imidazol-2-yl)propanoic acid (300 mg,1.579 mmol) in DCM (10 mL) was added HATU (722 mg, 1.895 mmol) and TEA(478 mg, 4.737 mmol). After stirring for 30 min at room temperature,(R)-1-amino-3-(3,4-dihydro isoquinolin-2(1H)-yl)propan-2-ol (488 mg,2.368 mmol) was added and the resulting mixture then stirred at roomtemperature for 16 h. Once TLC analysis showed the reaction to becomplete, the solvent was evaporated at reduced pressure and the residuepurified by preparative HPLC to give the title compound (184.6 mg,30.9%) as white solid. ¹H NMR (400 MHz, METHANOL-d₄) δ=7.56-7.46 (m,2H), 7.22-7.17 (m, 2H), 7.13-7.07 (m, 3H), 7.02-6.97 (m, 1H), 3.94 (t,J=5.8 Hz, 1H), 3.61 (d, J=3.3 Hz, 2H), 3.40-3.35 (m, 1H), 3.25-3.18 (m,3H), 2.89-2.84 (m, 2H), 2.79 (t, J=7.4 Hz, 2H), 2.74-2.69 (m, 2H), 2.47(d, J=6.5 Hz, 2H). LCMS (m/z): 379.1 [M+H]⁺.

Biological Assays PRMT5 Biochemical Assay

General Materials. S-adenosylmethionine (SAM), S-adenosylhomocysteine(SAH), bicine, KCl, Tween20, dimethylsulfoxide (DMSO), bovine skingelatin (BSG), and Tris(2-carboxyethyl)phosphine hydrochloride solution(TCEP) were purchased from Sigma-Aldrich at the highest level of puritypossible. ³H-SAM was purchase from American Radiolabeled Chemicals witha specific activity of 80 Ci/mmol. 384-well streptavidin Flashplateswere purchased from PerkinElmer.

Substrates. Peptide representative of human histone H4 residues 1-15 wassynthesized with a C-terminal linker-affinity tag motif and a C-terminalamide cap by 21^(st) Century Biochemicals. The peptide was highhigh-performance liquid chromatography (HPLC) purified to greater than95% purity and confirmed by liquid chromatography mass spectrometry(LC-MS). The sequence was Ac-SGRGKGGKGLGKGGA[K-Biot]-amide (SEQ ID NO.:3).

Molecular Biology: Full-length human PRMT5 (NM_(—)006109.3) transcriptvariant 1 clone was amplified from a fetal brain cDNA library,incorporating flanking 5′ sequence encoding a FLAG tag (MDYKDDDDK) (SEQID NO.: 4) fused directly to Ala 2 of PRMT5. Full-length human MEP50(NM_(—)024102) clone was amplified from a human testis cDNA libraryincorporating a 5′ sequence encoding a 6-histidine tag (MHHHHHH) (SEQ IDNO.: 5) fused directly to Arg 2 of MEP50. The amplified genes weresublconed into pENTR/D/TEV (Life Technologies) and subsequentlytransferred by Gateway™ attL×attR recombination to pDEST8 baculvirusexpression vector (Life Technologies).

Protein Expression. Recombinant baculovirus and Baculovirus-InfectedInsect Cells (BIIC) were generated according to Bac-to-Bac kitinstructions (Life Technologies) and Wasilko, 2006, respectively.Protein over-expression was accomplished by infecting exponentiallygrowing Spodoptera frugiperda (SF9) cell culture at 1.2×10⁶ cell/ml witha 5000 fold dilution of BIIC stock. Infections were carried out at 27°C. for 72 hours, harvested by centrifugation, and stored at −80° C. forpurification.

Protein Purification. Expressed full-length human Flag-PRMT5/6His-MeP50protein complex was purified from cell paste by NiNTA agarose affinitychromatography after a five hour equilibration of the resin with buffercontaining 50 mM Tris-HCL, pH 8.0, 25 mM NaCl, and 1 mM TCEP at 4° C.,to minimize the adsorption of tubulin impurity by the resin.Flag-PRMT5/6H is-MeP50 was eluted with 300 mM Imidazole in the samebuffer. The purity of recovered protein was 87%. Reference: Wasilko, D.J. and S. E. Lee: “TIPS: titerless infected-cells preservation andscale-up” Bioprocess J., 5 (2006), pp. 29-32.

Predicted Translations:

Flag-PRMT5 (SEQ ID NO.: 6) MDYKDDDDKA AMAVGGAGGS RVSSGRDLNC VPEIADTLGAVAKQGFDFLC MPVFHPRFKR EFIQEPAKNR PGPQTRSDLLLSGRDWNTLI VGKLSPWIRP DSKVEKIRRN SEAAMLQELNFGAYLGLPAF LLPLNQEDNT NLARVLTNHI HTGHHSSMFWMRVPLVAPED LRDDIIENAP TTHTEEYSGE EKTWMWWHNFRTLCDYSKRI AVALEIGADL PSNHVIDRWL GEPIKAAILPTSIFLTNKKG FPVLSKMHQR LIFRLLKLEV QFIITGTNHHSEKEFCSYLQ YLEYLSQNRP PPNAYELFAK GYEDYLQSPLQPLMDNLESQ TYEVFEKDPI KYSQYQQAIY KCLLDRVPEEEKDTNVQVLM VLGAGRGPLV NASLRAAKQA DRRIKLYAVEKNPNAVVTLE NWQFEEWGSQ VTVVSSDMRE WVAPEKADIIVSELLGSFAD NELSPECLDG AQHFLKDDGV SIPGEYTSFLAPISSSKLYN EVRACREKDR DPEAQFEMPY VVRLHNFHQLSAPQPCFTFS HPNRDPMIDN NRYCTLEFPV EVNTVLHGFAGYFETVLYQD ITLSIRPETH SPGMFSWFPI LFPIKQPITVREGQTICVRF WRCSNSKKVW YEWAVTAPVC SAIHNPTGRS YTIG L 6His-MEP50(SEQ ID NO.: 7) MHHHHHHRKE TPPPLVPPAA REWNLPPNAP ACMERQLEAARYRSDGALLL GASSLSGRCW AGSLWLFKDP CAAPNEGFCSAGVQTEAGVA DLTWVGERGI LVASDSGAVE LWELDENETLIVSKFCKYEH DDIVSTVSVL SSGTQAVSGS KDICIKVWDLAQQVVLSSYR AHAAQVTCVA ASPHKDSVFL SCSEDNRILLWDTRCPKPAS QIGCSAPGYL PTSLAWHPQQ SEVFVFGDENGTVSLVDTKS TSCVLSSAVH SQCVTGLVFS PHSVPFLASLSEDCSLAVLD SSLSELFRSQ AHRDFVRDAT WSPLNHSLLTTVGWDHQVVH HVVPTEPLPA PGPASVTE

General Procedure for PRMT5/MEP50 Enzyme Assays on Peptide Substrates.The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 ul) were spotted into a polypropylene384-well V-bottom plates (Greiner) using a Platemate Plus outfitted witha 384-channel head (Thermo Scientific). DMSO (1 ul) was added to Columns11, 12, 23, 24, rows A-H for the maximum signal control and 1 ul of SAH,a known product and inhibitor of PRMT5/MEP50, was added to columns 11,12, 23, 24, rows I-P for the minimum signal control. A cocktail (40 ul)containing the PRMT5/MEP50 enzyme and the peptide was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate withPRMT5/MEP50 for 30 min at 25 degrees Celsius, then a cocktail (10 ul)containing ³H-SAM was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows:PRMT5/MEP50 was 4 nM, ³H-SAM was 75 nM, peptide was 40 nM, SAH in theminimum signal control wells was 100 uM, and the DMSO concentration was1%. The assays were stopped by the addition of non-radioactive SAM (10ul) to a final concentration of 600 uM, which dilutes the ³H-SAM to alevel where its incorporation into the peptide substrate is no longerdetectable. 50 ul of the reaction in the 384-well polypropylene platewas then transferred to a 384-well Flashplate and the biotinylatedpeptides were allowed to bind to the streptavidin surface for at least 1hour before being washed three times with 0.1% Tween20 in a BiotekEL×405 plate washer. The plates were then read in a PerkinElmer TopCountplate reader to measure the quantity of ³H-labeled peptide bound to theFlashplate surface, measured as disintegrations per minute (dpm) oralternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {\left( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} \right) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{\left( {{Top} - {Bottom}} \right)}{\left( {1 + \left( \frac{X}{{IC}_{50}} \right)^{HillCoefficient}} \right.}}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

Z-138 Methylation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum, and D-PBSwere purchased from Life Technologies, Grand Island, N.Y., USA. Odysseyblocking buffer, 800CW goat anti-rabbit IgG (H+L) antibody, and LicorOdyssey infrared scanner were purchased from Licor Biosciences, Lincoln,Nebr., USA. Symmetric di-methyl arginine antibody was purchased from EMDMillipore, Billerica, Mass., USA. 16% Paraformaldehyde was purchasedfrom Electron Microscopy Sciences, Hatfield, Pa., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum and 100units/mL penicillin-streptomycin) and cultured at 37° C. under 5% CO₂.

Cell Treatment, In Cell Western (ICW) for detection of Symmetricdi-Methyl Arginine and DNA content. Z-138 cells were seeded in assaymedium at a concentration of 50,000 cells per mL to a 384-well cellculture plate with 50 μL per well. Compound (100 nL) from 384 wellsource plates was added directly to 384 well cell plate. Plates wereincubated at 37° C., 5% CO₂ for 96 hours. After four days of incubation,40 μL of cells from incubated plates were added to poly-D-lysine coated384 well culture plates (BD Biosciences 356697). Plates were incubatedat room temperature for 30 minutes then incubated at 37° C., 5% CO₂ for5 hours. After the incubation, 40 μL per well of 8% paraformaldehyde inPBS (16% paraformaldahyde was diluted to 8% in PBS) was added to eachplate and incubated for 30 minutes. Plates were transferred to a Biotek405 plate washer and washed 5 times with 100 μL per well of wash buffer(1×PBS with 0.1% Triton X-100 (v/v)). Next 30 μL per well of Odysseyblocking buffer were added to each plate and incubated 1 hour at roomtemperature. Blocking buffer was removed and 20 μL per well of primaryantibody was added (symmetric di-methyl arginine diluted 1:100 inOdyssey buffer with 0.1% Tween 20 (v/v)) and plates were incubatedovernight (16 hours) at 4° C. Plates were washed 5 times with 100 μL perwell of wash buffer. Next 20 μL per well of secondary antibody was added(1:200 800CW goat anti-rabbit IgG (H+L) antibody, 1:1000 DRAQ5(Biostatus limited) in Odyssey buffer with 0.1% Tween 20 (v/v)) andincubated for 1 hour at room temperature. The plates were washed 5 timeswith 100 μL per well wash buffer then 1 time with 100 μL per well ofwater. Plates were allowed to dry at room temperature then imaged on theLicor Odyssey machine which measures integrated intensity at 700 nm and800 nm wavelengths. Both 700 and 800 channels were scanned.

Calculations: First, the ratio for each well was determined by:

$\left( \frac{{symmetric}\mspace{14mu} {di}\text{-}{methyl}\mspace{14mu} {Arginine}\mspace{14mu} 800\mspace{14mu} {nm}\mspace{14mu} {value}}{{DRAQ}\; 5\mspace{14mu} 700\mspace{14mu} {nm}\mspace{14mu} {value}} \right)$

Each plate included fourteen control wells of DMSO only treatment(minimum inhibition) as well as fourteen control wells for maximuminhibition treated with 3 μM of a reference compound (Background wells).The average of the ratio values for each control type was calculated andused to determine the percent inhibition for each test well in theplate. Reference compound was serially diluted three-fold in DMSO for atotal of nine test concentrations, beginning at 3 μM. Percent inhibitionwas determined and IC₅₀ curves were generated using triplicate wells perconcentration of compound.

${{Percent}\mspace{14mu} {Inhibition}} = {100 - \left( {\left( \frac{\left( {{Individual}\mspace{14mu} {Test}\mspace{14mu} {Sample}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Avg}\mspace{14mu} {Ratio}} \right)}{\left( {{Minimum}\mspace{14mu} {Inhibition}\mspace{14mu} {Ratio}} \right) - \left( {{Background}\mspace{14mu} {Average}\mspace{14mu} {Ratio}} \right)} \right)*100} \right)}$

Z-138 Proliferation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum werepurchased from Life Technologies, Grand Island, N.Y., USA. V-bottompolypropylene 384-well plates were purchased from Greiner Bio-One,Monroe, N.C., USA. Cell culture 384-well white opaque plates werepurchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo® waspurchased from Promega Corporation, Madison, Wis., USA. SpectraMax M5plate reader was purchased from Molecular Devices LLC, Sunnyvale,Calif., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum andcultured at 37° C. under 5% CO₂ Under assay conditions, cells wereincubated in assay medium (RPMI 1640 supplemented with 10% v/v heatinactivated fetal bovine serum and 100 units/mL penicillin-streptomycin)at 37° C. under 5% CO₂.

For the assessment of the effect of compounds on the proliferation ofthe Z-138 cell line, exponentially growing cells were plated in 384-wellwhite opaque plates at a density of 10,000 cells/ml in a final volume of50 μl of assay medium. A compound source plate was prepared byperforming triplicate nine-point 3-fold serial dilutions in DMSO,beginning at 10 mM (final top concentration of compound in the assay was20 μM and the DMSO was 0.2%). A 100 mL aliquot from the compound stockplate was added to its respective well in the cell plate. The 100%inhibition control consisted of cells treated with 200 nM finalconcentration of staurosporine and the 0% inhibition control consistedof DMSO treated cells. After addition of compounds, assay plates wereincubated for 5 days at 37° C., 5% CO₂, relative humidity >90%. Cellviability was measured by quantitation of ATP present in the cellcultures, adding 35 μl of Cell Titer Glo® reagent to the cell plates.Luminescence was read in the SpectraMax M5 microplate reader. Theconcentration of compound inhibiting cell viability by 50% wasdetermined using a 4-parametric fit of the normalized dose responsecurves.

Results for certain compounds described herein are shown in Table 2.

TABLE 2 Biological Assay Results Cmpd No Biochemical IC₅₀ ICW EC₅₀Proliferation EC₅₀ 1 B B — 2 C — — 3 C — — 4 A — — 5 D — — 6 A A B 7 B BD 8 B B D 9 B D D 10 C — — 11 B B D 12 B B D 13 C — — 14 C — — 15 B B C16 B B D 17 B B D 18 C C ** 19 A B C 20 A B C 21 B B ** 22 A B C 23 A BC 24 B — — 25 B — — 26 A B C 27 A B C 28 A B C 29 B B ** 30 B B D 31 C BD 32 B B D 33 C — — 34 B B D 35 B B D 36 B B ** 37 A A C 38 A A C 39 A AB 40 A B C 41 C — — 42 B B ** 43 A B C 44 B B D 45 A A B 46 B B D 47 A BC 48 A B D 49 A B C 50 A A C 51 D — — 52 C — — 53 A B C 54 B B — 55 B —— 56 C — — 57 D — — 58 D — — 59 C — — 60 B C — 61 C — — 62 C — — 63 D —— 64 A B C 65 A B C 66 A B C 67 A A C 68 A B ** 69 B C — 70 A B ** 71 AB ** 72 C — — 73 A A B 74 A B C 75 A A C 76 A B C 77 A A C 78 B B — 79 AB C 80 A B D 81 A A B 82 A A C 83 B B D 84 A B C 85 C C — 86 A B D 87 C— — 88 A B D 89 B C — 90 A B D 91 A B C 92 A A C 93 A A C 94 A B D 95 AB D 96 A B D 97 B B C 98 A A C 99 A B C 100 A A C 101 A A C 102 A A C103 A B ** 104 B C ** 105 A B C 106 B B ** 107 A A C 108 A B D 109 A A B110 A A B 111 A A B 112 B B ** 113 B B D 114 B C ** 115 D — — 116 C C **117 B B C 118 B C ** 119 A B D 120 B C ** 121 C — — 122 B C ** 123 A B C124 C C ** 125 C — — 126 E — — 127 B C ** 128 E — — 129 B C ** 130 A B C131 C — — 132 C — — 133 * — — 134 B C ** 135 C — — 136 C — — 137 B C **138 B C — 139 * — — 140 C — — 141 C — — 142 B B ** 143 C — — 144 * — —145 C — — 146 A B — 147 * — — 148 * — — 149 A B — 150 B — — 151 B — —152 C — — 153 A B D 154 C — — 155 A A C 156 C — — 157 C — — 158 A B C159 A A C 160 B C ** 161 B C ** 162 C C ** 163 A B C 164 A A B 165 B B** 166 C — — 167 C — — 168 A A C 169 A A C 170 A B D 171 C — — 172 B B D173 C — — 174 B B ** 175 A A B 176 C — — 177 B B D 178 B B C 179 B B D180 C — — 181 A A — 182 B B — 183 B B — 184 C — — 185 B — — 186 C — —187 C — — 188 C — — 189 B — — 190 A — — 191 A — — 192 B — — 193 C — —For Table 2, “A” indicates an IC₅₀ or EC₅₀ < 0.100 μM, “B” indicates anIC₅₀ or EC₅₀ of 0.101-1.000 μM, “C” indicates an IC₅₀ or EC₅₀ of1.001-10.000 μM, “D” indicates an IC₅₀ or EC₅₀ of 10.001-50 μM, and “E”indicates an IC₅₀ or EC₅₀ > 50 μM. “—” indicates no data shown. “*”indicates an IC₅₀ or EC₅₀ > 10 μM. “**” indicates an IC₅₀ or EC₅₀ > 20μM.

Other Embodiments

The foregoing has been a description of certain non-limiting embodimentsof the invention. Those of ordinary skill in the art will appreciatethat various changes and modifications to this description may be madewithout departing from the spirit or scope of the present invention, asdefined in the following claims.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein

represents a single or double bond; R¹ is hydrogen, R^(z), or—C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆ alkyl; X is abond, —O—, —N(R)—, —CR⁴R⁵—, —O—CR⁴R⁵, —N(R)—CR⁴R⁵—, —O—CR⁴R⁵—O—,—N(R)—CR⁴R⁵—O, —N(R)—CR⁴R⁵—N(R)—, —O—CR⁴R⁵—N(R)—, —CR⁴R⁵—O—,—CR⁴R⁵—N(R)—, —O—CR⁴R⁵—CR⁶R⁷—, —N(R)—CR⁴R⁵—CR⁶R⁷—, —CR⁶R⁷—CR⁴R⁵—O—,—CR⁶R⁷—CR⁴R⁵—N(R)—, or —CR⁶R⁷—CR⁴R⁵—; each R is independently hydrogenor optionally substituted C₁₋₆ aliphatic; R² and R³ are independentlyselected from the group consisting of hydrogen, halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted phenyl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R² and R³ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring; R⁴ and R⁵ are independently selected from the groupconsisting of hydrogen, halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁴ and R⁵ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring; R⁶ and R⁷ are independently selected from the groupconsisting of hydrogen, halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedphenyl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or R⁶ and R⁷ are taken together withtheir intervening atoms to form an optionally substituted carbocyclic orheterocyclic ring; each R^(A) is independently selected from the groupconsisting of hydrogen, optionally substituted aliphatic, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, and optionally substituted heteroaryl; each R^(B) isindependently selected from the group consisting of hydrogen, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, and optionallysubstituted heteroaryl, or two R^(B) groups are taken together withtheir intervening atoms to form an optionally substituted heterocyclicring; R⁸, R⁹, R¹⁰, and R¹¹ are independently hydrogen, halo, oroptionally substituted aliphatic; Cy is a monocyclic or bicyclic,saturated, partially unsaturated, or aromatic ring having 0-4heteroatoms independently selected from nitrogen, oxygen, and sulfur,wherein Cy is substituted with 0, 1, 2, 3, or 4 R^(y) groups; each R^(y)is independently selected from the group consisting of halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; or an R^(y) group may be optionallytaken together with R² or R³ to form an optionally substituted 5- to6-membered carbocyclic or heterocyclic ring fused to Cy; each R^(x) isindependently selected from the group consisting of halo, —CN,optionally substituted aliphatic, —OR′, and —N(R″)₂; R′ is hydrogen oroptionally substituted aliphatic; each R″ is independently hydrogen oroptionally substituted aliphatic, or two R″ are taken together withtheir intervening atoms to form an optionally substituted heterocyclicring having 1-2 heteroatoms independently selected from nitrogen,oxygen, and sulfur; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, asvalency permits. 2-6. (canceled)
 7. The compound of claim 1, wherein thecompound is of Formula (II):

or a pharmaceutically acceptable salt thereof. 8-9. (canceled)
 10. Thecompound of claim 1, wherein the compound is of Formula (III):

or a pharmaceutically acceptable salt thereof. 11-12. (canceled)
 13. Thecompound of claim 1, wherein the compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof. 14-18. (canceled)
 19. Thecompound of claim 1, wherein R¹ is hydrogen.
 20. The compound of claim1, wherein n is 0, 1, or
 2. 21-23. (canceled)
 24. The compound of claim1, wherein R² is hydrogen. 25-30. (canceled)
 31. The compound of claim1, wherein R is hydrogen.
 32. (canceled)
 33. The compound of claim 1,wherein Cy is phenyl substituted with 0, 1, 2, 3, or 4 R^(y) groups.34-35. (canceled)
 36. The compound of claim 1, wherein Cy is a 5- to6-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and is substituted with 0, 1, 2, 3, or 4R^(y) groups. 37-39. (canceled)
 40. The compound of claim 1, wherein Cyis a bicyclic saturated, partially unsaturated, or aromatic ring having0-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur, wherein Cy is substituted with 0, 1, 2, 3, or 4 R^(y) groups.41-99. (canceled)
 100. The compound of claim 1, wherein the compound isselected from the group consisting of:


101. A pharmaceutical composition comprising a compound of claim 1 or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.
 102. A kit or packaged pharmaceutical comprising acompound of claim 1 or a pharmaceutically acceptable salt thereof, andinstructions for use thereof. 103-107. (canceled)
 108. A method oftreating a PRMT5-mediated disorder, comprising administering to asubject in need thereof a therapeutically effective amount of a compoundof claim 1 or a pharmaceutically acceptable salt thereof. 109.(canceled)
 110. The method of claim 108, wherein the disorder is cancer,a metabolic disorder, or a blood disorder.
 111. The method of claim 110,wherein the cancer is hematopoietic cancer, lung cancer, prostatecancer, melanoma, or pancreatic cancer.
 112. (canceled)
 113. The methodof claim 110, wherein the metabolic disorder is diabetes or obesity.114-115. (canceled)
 116. The method of claim 110, wherein the blooddisorder is a hemoglobinopathy.
 117. The method of claim 116, whereinthe blood disorder is sickle cell anemia or β-thalessemia. 118.(canceled)